Toolkit/directed evolution

directed evolution

Engineering Method·Research·Since 2018

Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.

Summary

Directed evolution is an engineering method that improves biological tool performance by iteratively selecting functional protein variants. In the cited split fluorescent protein study, it was demonstrated as one of two approaches used to improve split fluorescent proteins, contributing to brighter split sfCherry3 variants.

Usefulness & Problems

Why this is useful

Directed evolution is useful for enhancing protein function when rational prediction alone is insufficient. In the provided evidence, it enabled improvement of split fluorescent proteins that support endogenous protein tagging by gene editing and multiplexed visualization applications in living Caenorhabditis elegans.

Source:

for multiplexed visualization of neuronal synapses in living C. elegans

Source:

facilitating the tagging of endogenous proteins by gene editing

Source:

Based on sfCherry3, we have further developed a new red-colored trans-synaptic marker called Neuroligin-1 sfCherry3 Linker Across Synaptic Partners (NLG-1 CLASP)

Source:

CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as gene therapy, gene regulation, modifying epigenomes, and imaging chromosomes.

Problem solved

This method addresses the problem of suboptimal performance in engineered protein tools by enabling selection of improved variants from sequence diversity. In the cited context, it helped solve limited split fluorescent protein performance by improving split FP function and brightness.

Source:

for multiplexed visualization of neuronal synapses in living C. elegans

Source:

facilitating the tagging of endogenous proteins by gene editing

Source:

Based on sfCherry3, we have further developed a new red-colored trans-synaptic marker called Neuroligin-1 sfCherry3 Linker Across Synaptic Partners (NLG-1 CLASP)

Source:

CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as gene therapy, gene regulation, modifying epigenomes, and imaging chromosomes.

Taxonomy & Function

Primary hierarchy

Technique Branch

Method: A concrete method used to build, optimize, or evolve an engineered system.

Target processes

No target processes tagged yet.

Implementation Constraints

cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: builderswitch architecture: split

The cited implementation context is protein engineering of split fluorescent proteins, where directed evolution was used alongside SpyTag/SpyCatcher-assisted complementation as an improvement strategy. The evidence does not provide construct design details, host systems for selection, library generation methods, or screening assay parameters.

The supplied evidence does not specify the mutagenesis scheme, screening workflow, number of rounds, or quantitative gains attributable specifically to directed evolution alone. Independent replication and performance across multiple tool classes are not established by the provided sources.

Validation

Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication

Supporting Sources

Ranked Claims

Claim 1application demosupports2019Source 1needs review

NLG-1 CLASP enables multiplexed visualization of neuronal synapses in living C. elegans.

for multiplexed visualization of neuronal synapses in living C. elegans
Claim 2application demosupports2019Source 1needs review

NLG-1 CLASP enables multiplexed visualization of neuronal synapses in living C. elegans.

for multiplexed visualization of neuronal synapses in living C. elegans
Claim 3application demosupports2019Source 1needs review

NLG-1 CLASP enables multiplexed visualization of neuronal synapses in living C. elegans.

for multiplexed visualization of neuronal synapses in living C. elegans
Claim 4application demosupports2019Source 1needs review

NLG-1 CLASP enables multiplexed visualization of neuronal synapses in living C. elegans.

for multiplexed visualization of neuronal synapses in living C. elegans
Claim 5application demosupports2019Source 1needs review

NLG-1 CLASP enables multiplexed visualization of neuronal synapses in living C. elegans.

for multiplexed visualization of neuronal synapses in living C. elegans
Claim 6application demosupports2019Source 1needs review

NLG-1 CLASP enables multiplexed visualization of neuronal synapses in living C. elegans.

for multiplexed visualization of neuronal synapses in living C. elegans
Claim 7application demosupports2019Source 1needs review

NLG-1 CLASP enables multiplexed visualization of neuronal synapses in living C. elegans.

for multiplexed visualization of neuronal synapses in living C. elegans
Claim 8application demosupports2019Source 1needs review

NLG-1 CLASP enables multiplexed visualization of neuronal synapses in living C. elegans.

for multiplexed visualization of neuronal synapses in living C. elegans
Claim 9application demosupports2019Source 1needs review

NLG-1 CLASP enables multiplexed visualization of neuronal synapses in living C. elegans.

for multiplexed visualization of neuronal synapses in living C. elegans
Claim 10application demosupports2019Source 1needs review

NLG-1 CLASP enables multiplexed visualization of neuronal synapses in living C. elegans.

for multiplexed visualization of neuronal synapses in living C. elegans
Claim 11application enablementsupports2019Source 1needs review

The split sfCherry3 variants facilitate tagging of endogenous proteins by gene editing.

facilitating the tagging of endogenous proteins by gene editing
Claim 12application enablementsupports2019Source 1needs review

The split sfCherry3 variants facilitate tagging of endogenous proteins by gene editing.

facilitating the tagging of endogenous proteins by gene editing
Claim 13application enablementsupports2019Source 1needs review

The split sfCherry3 variants facilitate tagging of endogenous proteins by gene editing.

facilitating the tagging of endogenous proteins by gene editing
Claim 14application enablementsupports2019Source 1needs review

The split sfCherry3 variants facilitate tagging of endogenous proteins by gene editing.

facilitating the tagging of endogenous proteins by gene editing
Claim 15application enablementsupports2019Source 1needs review

The split sfCherry3 variants facilitate tagging of endogenous proteins by gene editing.

facilitating the tagging of endogenous proteins by gene editing
Claim 16application enablementsupports2019Source 1needs review

The split sfCherry3 variants facilitate tagging of endogenous proteins by gene editing.

facilitating the tagging of endogenous proteins by gene editing
Claim 17application enablementsupports2019Source 1needs review

The split sfCherry3 variants facilitate tagging of endogenous proteins by gene editing.

facilitating the tagging of endogenous proteins by gene editing
Claim 18application enablementsupports2019Source 1needs review

The split sfCherry3 variants facilitate tagging of endogenous proteins by gene editing.

facilitating the tagging of endogenous proteins by gene editing
Claim 19application enablementsupports2019Source 1needs review

The split sfCherry3 variants facilitate tagging of endogenous proteins by gene editing.

facilitating the tagging of endogenous proteins by gene editing
Claim 20application enablementsupports2019Source 1needs review

The split sfCherry3 variants facilitate tagging of endogenous proteins by gene editing.

facilitating the tagging of endogenous proteins by gene editing
Claim 21method improvementsupports2019Source 1needs review

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution
Claim 22method improvementsupports2019Source 1needs review

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution
Claim 23method improvementsupports2019Source 1needs review

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution
Claim 24method improvementsupports2019Source 1needs review

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution
Claim 25method improvementsupports2019Source 1needs review

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution
Claim 26method improvementsupports2019Source 1needs review

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution
Claim 27method improvementsupports2019Source 1needs review

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution
Claim 28method improvementsupports2019Source 1needs review

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution
Claim 29method improvementsupports2019Source 1needs review

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution
Claim 30method improvementsupports2019Source 1needs review

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution
Claim 31method improvementsupports2019Source 1needs review

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution
Claim 32method improvementsupports2019Source 1needs review

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution
Claim 33method improvementsupports2019Source 1needs review

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution
Claim 34method improvementsupports2019Source 1needs review

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution
Claim 35method improvementsupports2019Source 1needs review

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution
Claim 36method improvementsupports2019Source 1needs review

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution
Claim 37method improvementsupports2019Source 1needs review

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution
Claim 38performance improvementsupports2019Source 1needs review

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness
Claim 39performance improvementsupports2019Source 1needs review

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness
Claim 40performance improvementsupports2019Source 1needs review

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness
Claim 41performance improvementsupports2019Source 1needs review

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness
Claim 42performance improvementsupports2019Source 1needs review

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness
Claim 43performance improvementsupports2019Source 1needs review

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness
Claim 44performance improvementsupports2019Source 1needs review

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness
Claim 45performance improvementsupports2019Source 1needs review

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness
Claim 46performance improvementsupports2019Source 1needs review

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness
Claim 47performance improvementsupports2019Source 1needs review

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness
Claim 48performance improvementsupports2019Source 1needs review

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness
Claim 49performance improvementsupports2019Source 1needs review

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness
Claim 50performance improvementsupports2019Source 1needs review

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness
Claim 51performance improvementsupports2019Source 1needs review

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness
Claim 52performance improvementsupports2019Source 1needs review

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness
Claim 53performance improvementsupports2019Source 1needs review

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness
Claim 54performance improvementsupports2019Source 1needs review

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness
Claim 55tool developmentsupports2019Source 1needs review

Based on sfCherry3, the authors developed a new red-colored trans-synaptic marker called NLG-1 CLASP.

Based on sfCherry3, we have further developed a new red-colored trans-synaptic marker called Neuroligin-1 sfCherry3 Linker Across Synaptic Partners (NLG-1 CLASP)
Claim 56tool developmentsupports2019Source 1needs review

Based on sfCherry3, the authors developed a new red-colored trans-synaptic marker called NLG-1 CLASP.

Based on sfCherry3, we have further developed a new red-colored trans-synaptic marker called Neuroligin-1 sfCherry3 Linker Across Synaptic Partners (NLG-1 CLASP)
Claim 57tool developmentsupports2019Source 1needs review

Based on sfCherry3, the authors developed a new red-colored trans-synaptic marker called NLG-1 CLASP.

Based on sfCherry3, we have further developed a new red-colored trans-synaptic marker called Neuroligin-1 sfCherry3 Linker Across Synaptic Partners (NLG-1 CLASP)
Claim 58tool developmentsupports2019Source 1needs review

Based on sfCherry3, the authors developed a new red-colored trans-synaptic marker called NLG-1 CLASP.

Based on sfCherry3, we have further developed a new red-colored trans-synaptic marker called Neuroligin-1 sfCherry3 Linker Across Synaptic Partners (NLG-1 CLASP)
Claim 59tool developmentsupports2019Source 1needs review

Based on sfCherry3, the authors developed a new red-colored trans-synaptic marker called NLG-1 CLASP.

Based on sfCherry3, we have further developed a new red-colored trans-synaptic marker called Neuroligin-1 sfCherry3 Linker Across Synaptic Partners (NLG-1 CLASP)
Claim 60tool developmentsupports2019Source 1needs review

Based on sfCherry3, the authors developed a new red-colored trans-synaptic marker called NLG-1 CLASP.

Based on sfCherry3, we have further developed a new red-colored trans-synaptic marker called Neuroligin-1 sfCherry3 Linker Across Synaptic Partners (NLG-1 CLASP)
Claim 61tool developmentsupports2019Source 1needs review

Based on sfCherry3, the authors developed a new red-colored trans-synaptic marker called NLG-1 CLASP.

Based on sfCherry3, we have further developed a new red-colored trans-synaptic marker called Neuroligin-1 sfCherry3 Linker Across Synaptic Partners (NLG-1 CLASP)
Claim 62tool developmentsupports2019Source 1needs review

Based on sfCherry3, the authors developed a new red-colored trans-synaptic marker called NLG-1 CLASP.

Based on sfCherry3, we have further developed a new red-colored trans-synaptic marker called Neuroligin-1 sfCherry3 Linker Across Synaptic Partners (NLG-1 CLASP)
Claim 63tool developmentsupports2019Source 1needs review

Based on sfCherry3, the authors developed a new red-colored trans-synaptic marker called NLG-1 CLASP.

Based on sfCherry3, we have further developed a new red-colored trans-synaptic marker called Neuroligin-1 sfCherry3 Linker Across Synaptic Partners (NLG-1 CLASP)
Claim 64tool developmentsupports2019Source 1needs review

Based on sfCherry3, the authors developed a new red-colored trans-synaptic marker called NLG-1 CLASP.

Based on sfCherry3, we have further developed a new red-colored trans-synaptic marker called Neuroligin-1 sfCherry3 Linker Across Synaptic Partners (NLG-1 CLASP)
Claim 65application scopesupports2018Source 2needs review

CRISPR-Cas9 has been used across applications including gene therapy, gene regulation, epigenome modification, and chromosome imaging.

CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as gene therapy, gene regulation, modifying epigenomes, and imaging chromosomes.
Claim 66application scopesupports2018Source 2needs review

CRISPR-Cas9 has been used across applications including gene therapy, gene regulation, epigenome modification, and chromosome imaging.

CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as gene therapy, gene regulation, modifying epigenomes, and imaging chromosomes.
Claim 67application scopesupports2018Source 2needs review

CRISPR-Cas9 has been used across applications including gene therapy, gene regulation, epigenome modification, and chromosome imaging.

CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as gene therapy, gene regulation, modifying epigenomes, and imaging chromosomes.
Claim 68application scopesupports2018Source 2needs review

CRISPR-Cas9 has been used across applications including gene therapy, gene regulation, epigenome modification, and chromosome imaging.

CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as gene therapy, gene regulation, modifying epigenomes, and imaging chromosomes.
Claim 69application scopesupports2018Source 2needs review

CRISPR-Cas9 has been used across applications including gene therapy, gene regulation, epigenome modification, and chromosome imaging.

CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as gene therapy, gene regulation, modifying epigenomes, and imaging chromosomes.
Claim 70application scopesupports2018Source 2needs review

CRISPR-Cas9 has been used across applications including gene therapy, gene regulation, epigenome modification, and chromosome imaging.

CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as gene therapy, gene regulation, modifying epigenomes, and imaging chromosomes.
Claim 71application scopesupports2018Source 2needs review

CRISPR-Cas9 has been used across applications including gene therapy, gene regulation, epigenome modification, and chromosome imaging.

CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as gene therapy, gene regulation, modifying epigenomes, and imaging chromosomes.
Claim 72application scopesupports2018Source 2needs review

CRISPR-Cas9 has been used across applications including gene therapy, gene regulation, epigenome modification, and chromosome imaging.

CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as gene therapy, gene regulation, modifying epigenomes, and imaging chromosomes.
Claim 73application scopesupports2018Source 2needs review

CRISPR-Cas9 has been used across applications including gene therapy, gene regulation, epigenome modification, and chromosome imaging.

CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as gene therapy, gene regulation, modifying epigenomes, and imaging chromosomes.
Claim 74application scopesupports2018Source 2needs review

CRISPR-Cas9 has been used across applications including gene therapy, gene regulation, epigenome modification, and chromosome imaging.

CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as gene therapy, gene regulation, modifying epigenomes, and imaging chromosomes.
Claim 75impactsupports2018Source 2needs review

These Cas9 limitations hinder the use of CRISPR for disease treatment and wider biotechnological applications.

These obstacles hinder the use of CRISPR for disease treatment and in wider biotechnological applications.
Claim 76impactsupports2018Source 2needs review

These Cas9 limitations hinder the use of CRISPR for disease treatment and wider biotechnological applications.

These obstacles hinder the use of CRISPR for disease treatment and in wider biotechnological applications.
Claim 77impactsupports2018Source 2needs review

These Cas9 limitations hinder the use of CRISPR for disease treatment and wider biotechnological applications.

These obstacles hinder the use of CRISPR for disease treatment and in wider biotechnological applications.
Claim 78impactsupports2018Source 2needs review

These Cas9 limitations hinder the use of CRISPR for disease treatment and wider biotechnological applications.

These obstacles hinder the use of CRISPR for disease treatment and in wider biotechnological applications.
Claim 79impactsupports2018Source 2needs review

These Cas9 limitations hinder the use of CRISPR for disease treatment and wider biotechnological applications.

These obstacles hinder the use of CRISPR for disease treatment and in wider biotechnological applications.
Claim 80impactsupports2018Source 2needs review

These Cas9 limitations hinder the use of CRISPR for disease treatment and wider biotechnological applications.

These obstacles hinder the use of CRISPR for disease treatment and in wider biotechnological applications.
Claim 81impactsupports2018Source 2needs review

These Cas9 limitations hinder the use of CRISPR for disease treatment and wider biotechnological applications.

These obstacles hinder the use of CRISPR for disease treatment and in wider biotechnological applications.
Claim 82impactsupports2018Source 2needs review

These Cas9 limitations hinder the use of CRISPR for disease treatment and wider biotechnological applications.

These obstacles hinder the use of CRISPR for disease treatment and in wider biotechnological applications.
Claim 83impactsupports2018Source 2needs review

These Cas9 limitations hinder the use of CRISPR for disease treatment and wider biotechnological applications.

These obstacles hinder the use of CRISPR for disease treatment and in wider biotechnological applications.
Claim 84impactsupports2018Source 2needs review

These Cas9 limitations hinder the use of CRISPR for disease treatment and wider biotechnological applications.

These obstacles hinder the use of CRISPR for disease treatment and in wider biotechnological applications.
Claim 85limitationsupports2018Source 2needs review

Cas9 can exhibit aberrant off-target activity.

(ii) aberrant off-target activity
Claim 86limitationsupports2018Source 2needs review

Cas9 can exhibit aberrant off-target activity.

(ii) aberrant off-target activity
Claim 87limitationsupports2018Source 2needs review

Cas9 can exhibit aberrant off-target activity.

(ii) aberrant off-target activity
Claim 88limitationsupports2018Source 2needs review

Cas9 can exhibit aberrant off-target activity.

(ii) aberrant off-target activity
Claim 89limitationsupports2018Source 2needs review

Cas9 can exhibit aberrant off-target activity.

(ii) aberrant off-target activity
Claim 90limitationsupports2018Source 2needs review

Cas9 can exhibit aberrant off-target activity.

(ii) aberrant off-target activity
Claim 91limitationsupports2018Source 2needs review

Cas9 can exhibit aberrant off-target activity.

(ii) aberrant off-target activity
Claim 92limitationsupports2018Source 2needs review

Cas9 can exhibit aberrant off-target activity.

(ii) aberrant off-target activity
Claim 93limitationsupports2018Source 2needs review

Cas9 can exhibit aberrant off-target activity.

(ii) aberrant off-target activity
Claim 94limitationsupports2018Source 2needs review

Cas9 can exhibit aberrant off-target activity.

(ii) aberrant off-target activity
Claim 95limitationsupports2018Source 2needs review

Cas9 has a strict dependence on a protospacer-adjacent motif sequence.

some limitations have also been reported, for instance (i) a strict dependence on a protospacer-adjacent motif (PAM) sequence
Claim 96limitationsupports2018Source 2needs review

Cas9 has a strict dependence on a protospacer-adjacent motif sequence.

some limitations have also been reported, for instance (i) a strict dependence on a protospacer-adjacent motif (PAM) sequence
Claim 97limitationsupports2018Source 2needs review

Cas9 has a strict dependence on a protospacer-adjacent motif sequence.

some limitations have also been reported, for instance (i) a strict dependence on a protospacer-adjacent motif (PAM) sequence
Claim 98limitationsupports2018Source 2needs review

Cas9 has a strict dependence on a protospacer-adjacent motif sequence.

some limitations have also been reported, for instance (i) a strict dependence on a protospacer-adjacent motif (PAM) sequence
Claim 99limitationsupports2018Source 2needs review

Cas9 has a strict dependence on a protospacer-adjacent motif sequence.

some limitations have also been reported, for instance (i) a strict dependence on a protospacer-adjacent motif (PAM) sequence
Claim 100limitationsupports2018Source 2needs review

Cas9 has a strict dependence on a protospacer-adjacent motif sequence.

some limitations have also been reported, for instance (i) a strict dependence on a protospacer-adjacent motif (PAM) sequence
Claim 101limitationsupports2018Source 2needs review

Cas9 has a strict dependence on a protospacer-adjacent motif sequence.

some limitations have also been reported, for instance (i) a strict dependence on a protospacer-adjacent motif (PAM) sequence
Claim 102limitationsupports2018Source 2needs review

Cas9 has a strict dependence on a protospacer-adjacent motif sequence.

some limitations have also been reported, for instance (i) a strict dependence on a protospacer-adjacent motif (PAM) sequence
Claim 103limitationsupports2018Source 2needs review

Cas9 has a strict dependence on a protospacer-adjacent motif sequence.

some limitations have also been reported, for instance (i) a strict dependence on a protospacer-adjacent motif (PAM) sequence
Claim 104limitationsupports2018Source 2needs review

Cas9 has a strict dependence on a protospacer-adjacent motif sequence.

some limitations have also been reported, for instance (i) a strict dependence on a protospacer-adjacent motif (PAM) sequence
Claim 105limitationsupports2018Source 2needs review

Cas9 lacks sufficient modulation of protein binding and endonuclease activity for precise spatiotemporal control.

(iv) lack of modulation of protein binding and endonuclease activity, which is crucial for precise spatiotemporal control of gene expression or genome editing
Claim 106limitationsupports2018Source 2needs review

Cas9 lacks sufficient modulation of protein binding and endonuclease activity for precise spatiotemporal control.

(iv) lack of modulation of protein binding and endonuclease activity, which is crucial for precise spatiotemporal control of gene expression or genome editing
Claim 107limitationsupports2018Source 2needs review

Cas9 lacks sufficient modulation of protein binding and endonuclease activity for precise spatiotemporal control.

(iv) lack of modulation of protein binding and endonuclease activity, which is crucial for precise spatiotemporal control of gene expression or genome editing
Claim 108limitationsupports2018Source 2needs review

Cas9 lacks sufficient modulation of protein binding and endonuclease activity for precise spatiotemporal control.

(iv) lack of modulation of protein binding and endonuclease activity, which is crucial for precise spatiotemporal control of gene expression or genome editing
Claim 109limitationsupports2018Source 2needs review

Cas9 lacks sufficient modulation of protein binding and endonuclease activity for precise spatiotemporal control.

(iv) lack of modulation of protein binding and endonuclease activity, which is crucial for precise spatiotemporal control of gene expression or genome editing
Claim 110limitationsupports2018Source 2needs review

Cas9 lacks sufficient modulation of protein binding and endonuclease activity for precise spatiotemporal control.

(iv) lack of modulation of protein binding and endonuclease activity, which is crucial for precise spatiotemporal control of gene expression or genome editing
Claim 111limitationsupports2018Source 2needs review

Cas9 lacks sufficient modulation of protein binding and endonuclease activity for precise spatiotemporal control.

(iv) lack of modulation of protein binding and endonuclease activity, which is crucial for precise spatiotemporal control of gene expression or genome editing
Claim 112limitationsupports2018Source 2needs review

Cas9 lacks sufficient modulation of protein binding and endonuclease activity for precise spatiotemporal control.

(iv) lack of modulation of protein binding and endonuclease activity, which is crucial for precise spatiotemporal control of gene expression or genome editing
Claim 113limitationsupports2018Source 2needs review

Cas9 lacks sufficient modulation of protein binding and endonuclease activity for precise spatiotemporal control.

(iv) lack of modulation of protein binding and endonuclease activity, which is crucial for precise spatiotemporal control of gene expression or genome editing
Claim 114limitationsupports2018Source 2needs review

Cas9 lacks sufficient modulation of protein binding and endonuclease activity for precise spatiotemporal control.

(iv) lack of modulation of protein binding and endonuclease activity, which is crucial for precise spatiotemporal control of gene expression or genome editing
Claim 115limitationsupports2018Source 2needs review

The large size of Cas9 creates problems for CRISPR delivery.

(iii) the large size of Cas9 is problematic for CRISPR delivery
Claim 116limitationsupports2018Source 2needs review

The large size of Cas9 creates problems for CRISPR delivery.

(iii) the large size of Cas9 is problematic for CRISPR delivery
Claim 117limitationsupports2018Source 2needs review

The large size of Cas9 creates problems for CRISPR delivery.

(iii) the large size of Cas9 is problematic for CRISPR delivery
Claim 118limitationsupports2018Source 2needs review

The large size of Cas9 creates problems for CRISPR delivery.

(iii) the large size of Cas9 is problematic for CRISPR delivery
Claim 119limitationsupports2018Source 2needs review

The large size of Cas9 creates problems for CRISPR delivery.

(iii) the large size of Cas9 is problematic for CRISPR delivery
Claim 120limitationsupports2018Source 2needs review

The large size of Cas9 creates problems for CRISPR delivery.

(iii) the large size of Cas9 is problematic for CRISPR delivery
Claim 121limitationsupports2018Source 2needs review

The large size of Cas9 creates problems for CRISPR delivery.

(iii) the large size of Cas9 is problematic for CRISPR delivery
Claim 122limitationsupports2018Source 2needs review

The large size of Cas9 creates problems for CRISPR delivery.

(iii) the large size of Cas9 is problematic for CRISPR delivery
Claim 123limitationsupports2018Source 2needs review

The large size of Cas9 creates problems for CRISPR delivery.

(iii) the large size of Cas9 is problematic for CRISPR delivery
Claim 124limitationsupports2018Source 2needs review

The large size of Cas9 creates problems for CRISPR delivery.

(iii) the large size of Cas9 is problematic for CRISPR delivery
Claim 125review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 126review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 127review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 128review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 129review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 130review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 131review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 132review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 133review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 134review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 135review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 136review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 137review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 138review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 139review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 140review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 141review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 142review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 143review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 144review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 145review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 146review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 147review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 148review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 149review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 150review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 151review focussupports2018Source 2needs review

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Claim 152strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 153strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 154strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 155strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 156strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 157strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 158strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 159strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 160strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 161strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 162strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 163strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 164strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 165strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 166strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 167strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 168strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 169strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 170strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 171strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 172strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 173strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 174strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 175strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 176strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 177strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Claim 178strategy overviewsupports2018Source 2needs review

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.

Approval Evidence

2 sources4 linked approval claimsfirst-pass slug directed-evolution
we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution

Source:

studies that improve or develop novel protein functions through ... directed evolution

Source:

method improvementsupports

SpyTag/SpyCatcher interaction and directed evolution were demonstrated as two approaches to improve split fluorescent proteins.

we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution

Source:

performance improvementsupports

Directed evolution yielded two split sfCherry3 variants with substantially enhanced overall brightness.

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness

Source:

review focussupports

The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.

Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.

Source:

strategy overviewsupports

Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.

Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.

Source:

Comparisons

Source-backed strengths

The evidence supports directed evolution as an experimentally demonstrated route for improving split fluorescent proteins. Its practical strength here is association with improved split sfCherry3 performance and downstream utility in endogenous protein tagging and synapse visualization.

Source:

The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness

Compared with CoTV

directed evolution and CoTV address a similar problem space.

Shared frame: same top-level item type

Strengths here: looks easier to implement in practice.

directed evolution and gene editing technology address a similar problem space.

Shared frame: same top-level item type

directed evolution and light-dependent protein (un)folding reactions address a similar problem space.

Shared frame: same top-level item type

Strengths here: looks easier to implement in practice.

Ranked Citations

  1. 1.
    StructuralSource 1Communications Biology2019Claim 8Claim 8Claim 10

    Extracted from this source document.

  2. 2.
    StructuralSource 2International Journal of Genomics2018Claim 74Claim 74Claim 74

    Seeded from load plan for claim cl7. Extracted from this source document.