Source 1review2021Frontiers in Genome Editing
Toolkit/synthetically engineered guide RNA
synthetically engineered guide RNA
Also known as: engineered gRNA, gRNA, guide RNA, synthetic gRNA
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Synthetically engineered guide RNAs are chemically modified or conjugated CRISPR guide RNAs used in mammalian cells to improve the performance and functional scope of CRISPR-Cas9 systems. Reported implementations increase guide stability and specificity, support donor DNA tethering to enhance homology-directed repair, and enable fluorescent genomic imaging.
Usefulness & Problems
Why this is useful
These engineered RNAs are useful because they expand guide RNA function beyond target recognition alone, improving CRISPR-Cas9 stability, safety, specificity, and versatility in mammalian cells. The cited applications specifically include enhanced homology-directed repair through donor proximity and highly specific nuclear staining for chromosomal and genomic imaging.
Problem solved
This tool helps address several limitations of standard CRISPR guide RNAs, including insufficient stability, suboptimal editing specificity, and limited functional modularity. It also addresses the challenge of inefficient homology-directed repair by conjugating donor DNA to the guide RNA to increase local donor availability at the edited site.
Problem links
Need conditional control of signaling activity
DerivedSynthetically engineered guide RNAs are chemically modified or functionally conjugated CRISPR guide RNAs used in mammalian cells to improve genome editing performance. Reported implementations increase guide stability, reduce off-target editing and inflammatory signaling, and can be adapted for donor tethering or fluorescent genomic imaging.
Need conditional protein clearance
DerivedSynthetically engineered guide RNAs are chemically modified or functionally conjugated CRISPR guide RNAs used in mammalian cells to improve genome editing performance. Reported implementations increase guide stability, reduce off-target editing and inflammatory signaling, and can be adapted for donor tethering or fluorescent genomic imaging.
Need conditional recombination or state switching
DerivedSynthetically engineered guide RNAs are chemically modified or functionally conjugated CRISPR guide RNAs used in mammalian cells to improve genome editing performance. Reported implementations increase guide stability, reduce off-target editing and inflammatory signaling, and can be adapted for donor tethering or fluorescent genomic imaging.
Need controllable genome or transcript editing
DerivedSynthetically engineered guide RNAs are chemically modified or functionally conjugated CRISPR guide RNAs used in mammalian cells to improve genome editing performance. Reported implementations increase guide stability, reduce off-target editing and inflammatory signaling, and can be adapted for donor tethering or fluorescent genomic imaging.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Component: A low-level RNA part used inside a larger architecture that realizes a mechanism.
Mechanisms
Degradationdonor tethering/proximity enhancementdonor tethering/proximity enhancementfluorescent labeling for imagingfluorescent labeling for imagingnuclease-resistance-mediated stabilizationnuclease-resistance-mediated stabilizationoff-target suppressionoff-target suppressionreduced inflammatory signalingreduced inflammatory signalingTechniques
No technique tags yet.
Target processes
degradationeditingrecombinationsignalingImplementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: regulator
The available evidence supports implementation through chemical modification of guide RNAs, molecular conjugation of exogenous donor DNA, and fluorophore labeling for imaging applications in mammalian cells. However, the supplied material does not specify sequence design rules, Cas protein compatibility beyond CRISPR-Cas9 context, delivery methods, or required cofactors.
The supplied evidence is drawn from a single 2021 review-style source and does not provide detailed quantitative performance benchmarks, standardized construct designs, or side-by-side comparisons across guide formats. The evidence provided also does not specify which chemical modifications, fluorophores, donor attachment chemistries, or mammalian cell contexts were validated.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Guide RNAs conjugated with exogenous donor DNA are described as improving homology-directed repair efficiency by ensuring donor proximity to the edited site.
through conjugation with exogenous donor DNA, engineered gRNAs have been shown to improve homology-directed repair (HDR) efficiency by ensuring donor proximity to the edited site
Guide RNAs conjugated with exogenous donor DNA are described as improving homology-directed repair efficiency by ensuring donor proximity to the edited site.
through conjugation with exogenous donor DNA, engineered gRNAs have been shown to improve homology-directed repair (HDR) efficiency by ensuring donor proximity to the edited site
Guide RNAs conjugated with exogenous donor DNA are described as improving homology-directed repair efficiency by ensuring donor proximity to the edited site.
through conjugation with exogenous donor DNA, engineered gRNAs have been shown to improve homology-directed repair (HDR) efficiency by ensuring donor proximity to the edited site
Guide RNAs conjugated with exogenous donor DNA are described as improving homology-directed repair efficiency by ensuring donor proximity to the edited site.
through conjugation with exogenous donor DNA, engineered gRNAs have been shown to improve homology-directed repair (HDR) efficiency by ensuring donor proximity to the edited site
Guide RNAs conjugated with exogenous donor DNA are described as improving homology-directed repair efficiency by ensuring donor proximity to the edited site.
through conjugation with exogenous donor DNA, engineered gRNAs have been shown to improve homology-directed repair (HDR) efficiency by ensuring donor proximity to the edited site
Guide RNAs conjugated with exogenous donor DNA are described as improving homology-directed repair efficiency by ensuring donor proximity to the edited site.
through conjugation with exogenous donor DNA, engineered gRNAs have been shown to improve homology-directed repair (HDR) efficiency by ensuring donor proximity to the edited site
Guide RNAs conjugated with exogenous donor DNA are described as improving homology-directed repair efficiency by ensuring donor proximity to the edited site.
through conjugation with exogenous donor DNA, engineered gRNAs have been shown to improve homology-directed repair (HDR) efficiency by ensuring donor proximity to the edited site
Guide RNAs conjugated with exogenous donor DNA are described as improving homology-directed repair efficiency by ensuring donor proximity to the edited site.
through conjugation with exogenous donor DNA, engineered gRNAs have been shown to improve homology-directed repair (HDR) efficiency by ensuring donor proximity to the edited site
Guide RNAs conjugated with exogenous donor DNA are described as improving homology-directed repair efficiency by ensuring donor proximity to the edited site.
through conjugation with exogenous donor DNA, engineered gRNAs have been shown to improve homology-directed repair (HDR) efficiency by ensuring donor proximity to the edited site
Guide RNAs conjugated with exogenous donor DNA are described as improving homology-directed repair efficiency by ensuring donor proximity to the edited site.
through conjugation with exogenous donor DNA, engineered gRNAs have been shown to improve homology-directed repair (HDR) efficiency by ensuring donor proximity to the edited site
Fluorescently labeled synthetic guide RNAs are described as enabling highly specific nuclear staining and imaging for mechanistic studies of chromosomal dynamics and genomic mapping.
synthetic gRNAs attached to fluorescent labels have been developed to enable highly specific nuclear staining and imaging, enabling mechanistic studies of chromosomal dynamics and genomic mapping
Fluorescently labeled synthetic guide RNAs are described as enabling highly specific nuclear staining and imaging for mechanistic studies of chromosomal dynamics and genomic mapping.
synthetic gRNAs attached to fluorescent labels have been developed to enable highly specific nuclear staining and imaging, enabling mechanistic studies of chromosomal dynamics and genomic mapping
Fluorescently labeled synthetic guide RNAs are described as enabling highly specific nuclear staining and imaging for mechanistic studies of chromosomal dynamics and genomic mapping.
synthetic gRNAs attached to fluorescent labels have been developed to enable highly specific nuclear staining and imaging, enabling mechanistic studies of chromosomal dynamics and genomic mapping
Fluorescently labeled synthetic guide RNAs are described as enabling highly specific nuclear staining and imaging for mechanistic studies of chromosomal dynamics and genomic mapping.
synthetic gRNAs attached to fluorescent labels have been developed to enable highly specific nuclear staining and imaging, enabling mechanistic studies of chromosomal dynamics and genomic mapping
Fluorescently labeled synthetic guide RNAs are described as enabling highly specific nuclear staining and imaging for mechanistic studies of chromosomal dynamics and genomic mapping.
synthetic gRNAs attached to fluorescent labels have been developed to enable highly specific nuclear staining and imaging, enabling mechanistic studies of chromosomal dynamics and genomic mapping
Fluorescently labeled synthetic guide RNAs are described as enabling highly specific nuclear staining and imaging for mechanistic studies of chromosomal dynamics and genomic mapping.
synthetic gRNAs attached to fluorescent labels have been developed to enable highly specific nuclear staining and imaging, enabling mechanistic studies of chromosomal dynamics and genomic mapping
Fluorescently labeled synthetic guide RNAs are described as enabling highly specific nuclear staining and imaging for mechanistic studies of chromosomal dynamics and genomic mapping.
synthetic gRNAs attached to fluorescent labels have been developed to enable highly specific nuclear staining and imaging, enabling mechanistic studies of chromosomal dynamics and genomic mapping
Fluorescently labeled synthetic guide RNAs are described as enabling highly specific nuclear staining and imaging for mechanistic studies of chromosomal dynamics and genomic mapping.
synthetic gRNAs attached to fluorescent labels have been developed to enable highly specific nuclear staining and imaging, enabling mechanistic studies of chromosomal dynamics and genomic mapping
Fluorescently labeled synthetic guide RNAs are described as enabling highly specific nuclear staining and imaging for mechanistic studies of chromosomal dynamics and genomic mapping.
synthetic gRNAs attached to fluorescent labels have been developed to enable highly specific nuclear staining and imaging, enabling mechanistic studies of chromosomal dynamics and genomic mapping
Fluorescently labeled synthetic guide RNAs are described as enabling highly specific nuclear staining and imaging for mechanistic studies of chromosomal dynamics and genomic mapping.
synthetic gRNAs attached to fluorescent labels have been developed to enable highly specific nuclear staining and imaging, enabling mechanistic studies of chromosomal dynamics and genomic mapping
Engineered guide RNAs are described as improving CRISPR-Cas9 stability, specificity, safety, and versatility.
gRNAs have been engineered to improve the CRISPR system's overall stability, specificity, safety, and versatility.
Engineered guide RNAs are described as improving CRISPR-Cas9 stability, specificity, safety, and versatility.
gRNAs have been engineered to improve the CRISPR system's overall stability, specificity, safety, and versatility.
Engineered guide RNAs are described as improving CRISPR-Cas9 stability, specificity, safety, and versatility.
gRNAs have been engineered to improve the CRISPR system's overall stability, specificity, safety, and versatility.
Engineered guide RNAs are described as improving CRISPR-Cas9 stability, specificity, safety, and versatility.
gRNAs have been engineered to improve the CRISPR system's overall stability, specificity, safety, and versatility.
Engineered guide RNAs are described as improving CRISPR-Cas9 stability, specificity, safety, and versatility.
gRNAs have been engineered to improve the CRISPR system's overall stability, specificity, safety, and versatility.
Engineered guide RNAs are described as improving CRISPR-Cas9 stability, specificity, safety, and versatility.
gRNAs have been engineered to improve the CRISPR system's overall stability, specificity, safety, and versatility.
Synthetic guide RNA is described as reducing inflammatory signaling associated with the CRISPR system, thereby limiting immunogenicity and toxicity in edited mammalian cells.
Synthetic gRNA has been shown to ameliorate inflammatory signaling caused by the CRISPR system, thereby limiting immunogenicity and toxicity in edited mammalian cells.
Synthetic guide RNA is described as reducing inflammatory signaling associated with the CRISPR system, thereby limiting immunogenicity and toxicity in edited mammalian cells.
Synthetic gRNA has been shown to ameliorate inflammatory signaling caused by the CRISPR system, thereby limiting immunogenicity and toxicity in edited mammalian cells.
Synthetic guide RNA is described as reducing inflammatory signaling associated with the CRISPR system, thereby limiting immunogenicity and toxicity in edited mammalian cells.
Synthetic gRNA has been shown to ameliorate inflammatory signaling caused by the CRISPR system, thereby limiting immunogenicity and toxicity in edited mammalian cells.
Synthetic guide RNA is described as reducing inflammatory signaling associated with the CRISPR system, thereby limiting immunogenicity and toxicity in edited mammalian cells.
Synthetic gRNA has been shown to ameliorate inflammatory signaling caused by the CRISPR system, thereby limiting immunogenicity and toxicity in edited mammalian cells.
Synthetic guide RNA is described as reducing inflammatory signaling associated with the CRISPR system, thereby limiting immunogenicity and toxicity in edited mammalian cells.
Synthetic gRNA has been shown to ameliorate inflammatory signaling caused by the CRISPR system, thereby limiting immunogenicity and toxicity in edited mammalian cells.
Synthetic guide RNA is described as reducing inflammatory signaling associated with the CRISPR system, thereby limiting immunogenicity and toxicity in edited mammalian cells.
Synthetic gRNA has been shown to ameliorate inflammatory signaling caused by the CRISPR system, thereby limiting immunogenicity and toxicity in edited mammalian cells.
Engineered guide RNAs are described as improving guide specificity by limiting off-target editing.
guide specificity has been improved by limiting off-target editing
Engineered guide RNAs are described as improving guide specificity by limiting off-target editing.
guide specificity has been improved by limiting off-target editing
Engineered guide RNAs are described as improving guide specificity by limiting off-target editing.
guide specificity has been improved by limiting off-target editing
Engineered guide RNAs are described as improving guide specificity by limiting off-target editing.
guide specificity has been improved by limiting off-target editing
Engineered guide RNAs are described as improving guide specificity by limiting off-target editing.
guide specificity has been improved by limiting off-target editing
Engineered guide RNAs are described as improving guide specificity by limiting off-target editing.
guide specificity has been improved by limiting off-target editing
Guide RNA modifications are described as increasing guide stability against nuclease degradation and enhancing efficiency.
gRNAs have been modified to increase their stability to guard against nuclease degradation, thereby enhancing their efficiency.
Guide RNA modifications are described as increasing guide stability against nuclease degradation and enhancing efficiency.
gRNAs have been modified to increase their stability to guard against nuclease degradation, thereby enhancing their efficiency.
Guide RNA modifications are described as increasing guide stability against nuclease degradation and enhancing efficiency.
gRNAs have been modified to increase their stability to guard against nuclease degradation, thereby enhancing their efficiency.
Guide RNA modifications are described as increasing guide stability against nuclease degradation and enhancing efficiency.
gRNAs have been modified to increase their stability to guard against nuclease degradation, thereby enhancing their efficiency.
Guide RNA modifications are described as increasing guide stability against nuclease degradation and enhancing efficiency.
gRNAs have been modified to increase their stability to guard against nuclease degradation, thereby enhancing their efficiency.
Guide RNA modifications are described as increasing guide stability against nuclease degradation and enhancing efficiency.
gRNAs have been modified to increase their stability to guard against nuclease degradation, thereby enhancing their efficiency.
Approval Evidence
1 source4 linked approval claimsfirst-pass slug synthetically-engineered-guide-rna
Using Synthetically Engineered Guide RNAs to Enhance CRISPR Genome Editing Systems in Mammalian Cells
Source:
review summarysupports
Engineered guide RNAs are described as improving CRISPR-Cas9 stability, specificity, safety, and versatility.
gRNAs have been engineered to improve the CRISPR system's overall stability, specificity, safety, and versatility.
Source:
safety effectsupports
Synthetic guide RNA is described as reducing inflammatory signaling associated with the CRISPR system, thereby limiting immunogenicity and toxicity in edited mammalian cells.
Synthetic gRNA has been shown to ameliorate inflammatory signaling caused by the CRISPR system, thereby limiting immunogenicity and toxicity in edited mammalian cells.
Source:
specificity effectsupports
Engineered guide RNAs are described as improving guide specificity by limiting off-target editing.
guide specificity has been improved by limiting off-target editing
Source:
stability effectsupports
Guide RNA modifications are described as increasing guide stability against nuclease degradation and enhancing efficiency.
gRNAs have been modified to increase their stability to guard against nuclease degradation, thereby enhancing their efficiency.
Source:
Comparisons
Source-backed strengths
Reported strengths include improved CRISPR-Cas9 stability, specificity, safety, and versatility. Fluorescently labeled synthetic guide RNAs have been described as enabling highly specific nuclear staining and imaging for mechanistic studies of chromosomal dynamics and genomic mapping, and donor-conjugated guides have been described as improving homology-directed repair efficiency.
Compared with MALAT1
synthetically engineered guide RNA and MALAT1 address a similar problem space because they share degradation, recombination.
Shared frame: same top-level item type; shared target processes: degradation, recombination; shared mechanisms: degradation
Compared with PCB synthesis expression vector
synthetically engineered guide RNA and PCB synthesis expression vector address a similar problem space because they share degradation, recombination, signaling.
Shared frame: shared target processes: degradation, recombination, signaling; shared mechanisms: degradation
Strengths here: looks easier to implement in practice; may avoid an exogenous cofactor requirement.
Compared with Split-Cas9-based targeted gene editing
synthetically engineered guide RNA and Split-Cas9-based targeted gene editing address a similar problem space because they share degradation, editing.
Shared frame: shared target processes: degradation, editing; shared mechanisms: degradation
Strengths here: looks easier to implement in practice.
Ranked Citations
- 1.