Source 1primary paper2017Genome biology
Toolkit/eSpCas9
eSpCas9
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
eSpCas9 is an increased-fidelity Streptococcus pyogenes Cas9 nuclease variant used in the optimization of CRISPR-Cas9 cleavage specificity. In comparative analyses of high-fidelity SpCas9 enzymes, eSpCas9 served as one of the variants whose mutations were combined to generate hybrid HeFSpCas9 nucleases.
Usefulness & Problems
Why this is useful
eSpCas9 is useful as a high-fidelity SpCas9 option in workflows that require improved cleavage specificity relative to standard CRISPR-Cas9 editing. The cited comparative study indicates that highest-specificity cleavage is achieved by matching each target sequence with an appropriate high-fidelity nuclease rather than relying on a single universally best variant.
Source:
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
Problem solved
eSpCas9 helps address the problem of optimizing CRISPR-Cas9 target cleavage specificity across different genomic targets. The source specifically supports that no single high-fidelity SpCas9 variant is generally superior across targets, motivating target-by-target nuclease selection.
Source:
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
Problem links
Need conditional recombination or state switching
DerivedeSpCas9 is an increased-fidelity Streptococcus pyogenes Cas9 nuclease variant discussed in the context of optimizing CRISPR-Cas9 cleavage specificity. In comparative analyses with other high-fidelity SpCas9 variants, it contributed mutations used to generate HeFSpCas9 hybrid variants.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Component: A low-level protein part used inside a larger architecture that realizes a mechanism.
Techniques
No technique tags yet.
Target processes
recombinationImplementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: actuatoroperating role: regulatorswitch architecture: cleavage
For increased-fidelity nucleases including eSpCas9, a matching 5' G extension is reported to be more detrimental to activity than a mismatching 5' G extension. Beyond this guide-design consideration, the supplied evidence does not specify construct architecture, delivery modality, cofactor requirements, or expression system details.
The supplied evidence does not provide direct quantitative performance data for eSpCas9, such as on-target activity, off-target profiles, PAM constraints, or organism-specific validation. The cited study also concludes that no single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets, which limits claims of universal advantage.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
The paper provides a framework for generating new nuclease variants for targets that currently lack a matching optimal nuclease and a simple means for identifying the optimal nuclease when accurate target-ranking prediction tools are absent.
We provide here a framework for generating new nuclease variants for targets that currently have no matching optimal nuclease, and offer a simple mean for identifying the optimal nuclease for targets in the absence of accurate target-ranking prediction tools
The paper provides a framework for generating new nuclease variants for targets that currently lack a matching optimal nuclease and a simple means for identifying the optimal nuclease when accurate target-ranking prediction tools are absent.
We provide here a framework for generating new nuclease variants for targets that currently have no matching optimal nuclease, and offer a simple mean for identifying the optimal nuclease for targets in the absence of accurate target-ranking prediction tools
The paper provides a framework for generating new nuclease variants for targets that currently lack a matching optimal nuclease and a simple means for identifying the optimal nuclease when accurate target-ranking prediction tools are absent.
We provide here a framework for generating new nuclease variants for targets that currently have no matching optimal nuclease, and offer a simple mean for identifying the optimal nuclease for targets in the absence of accurate target-ranking prediction tools
The paper provides a framework for generating new nuclease variants for targets that currently lack a matching optimal nuclease and a simple means for identifying the optimal nuclease when accurate target-ranking prediction tools are absent.
We provide here a framework for generating new nuclease variants for targets that currently have no matching optimal nuclease, and offer a simple mean for identifying the optimal nuclease for targets in the absence of accurate target-ranking prediction tools
The paper provides a framework for generating new nuclease variants for targets that currently lack a matching optimal nuclease and a simple means for identifying the optimal nuclease when accurate target-ranking prediction tools are absent.
We provide here a framework for generating new nuclease variants for targets that currently have no matching optimal nuclease, and offer a simple mean for identifying the optimal nuclease for targets in the absence of accurate target-ranking prediction tools
The paper provides a framework for generating new nuclease variants for targets that currently lack a matching optimal nuclease and a simple means for identifying the optimal nuclease when accurate target-ranking prediction tools are absent.
We provide here a framework for generating new nuclease variants for targets that currently have no matching optimal nuclease, and offer a simple mean for identifying the optimal nuclease for targets in the absence of accurate target-ranking prediction tools
The paper provides a framework for generating new nuclease variants for targets that currently lack a matching optimal nuclease and a simple means for identifying the optimal nuclease when accurate target-ranking prediction tools are absent.
We provide here a framework for generating new nuclease variants for targets that currently have no matching optimal nuclease, and offer a simple mean for identifying the optimal nuclease for targets in the absence of accurate target-ranking prediction tools
The paper provides a framework for generating new nuclease variants for targets that currently lack a matching optimal nuclease and a simple means for identifying the optimal nuclease when accurate target-ranking prediction tools are absent.
We provide here a framework for generating new nuclease variants for targets that currently have no matching optimal nuclease, and offer a simple mean for identifying the optimal nuclease for targets in the absence of accurate target-ranking prediction tools
The paper provides a framework for generating new nuclease variants for targets that currently lack a matching optimal nuclease and a simple means for identifying the optimal nuclease when accurate target-ranking prediction tools are absent.
We provide here a framework for generating new nuclease variants for targets that currently have no matching optimal nuclease, and offer a simple mean for identifying the optimal nuclease for targets in the absence of accurate target-ranking prediction tools
The paper provides a framework for generating new nuclease variants for targets that currently lack a matching optimal nuclease and a simple means for identifying the optimal nuclease when accurate target-ranking prediction tools are absent.
We provide here a framework for generating new nuclease variants for targets that currently have no matching optimal nuclease, and offer a simple mean for identifying the optimal nuclease for targets in the absence of accurate target-ranking prediction tools
The paper concerns crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage.
The paper concerns crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage.
The paper concerns crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage.
The paper concerns crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage.
The paper concerns crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage.
The paper concerns crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage.
The paper concerns crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage.
The paper concerns crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage.
The paper concerns crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage.
The paper concerns crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage.
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
Approval Evidence
1 source8 linked approval claimsfirst-pass slug espcas9
eSpCas9
Source:
activity effectsupports
For the increased-fidelity nucleases, a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension.
a matching 5' G extension being more detrimental to their activities than a mismatching one
Source:
application guidancesupports
For highest-specificity cleavage, each target should be matched with an appropriate high-fidelity nuclease.
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
Source:
comparative conclusionsupports
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets.
No single nuclease variant shows generally superior fidelity
Source:
engineering resultsupports
HeFSpCas9 variants were generated by combining mutations from eSpCas9 and SpCas9-HF1.
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
Source:
mechanistic effectsupports
Mutations in the increased-fidelity SpCas9 variants may reduce cleavage without reducing DNA binding.
the mutations in these variants may diminish the cleavage, but not the DNA-binding, of SpCas9s
Source:
specificity improvementsupports
HeFSpCas9 variants show substantially improved specificity for targets where eSpCas9 and SpCas9-HF1 have higher off-target propensity.
HeFSpCas9s exhibit substantially improved specificity specifically for those targets for which eSpCas9 and SpCas9-HF1 have higher off-target propensity
Source:
target dependencesupports
Targets can be ranked by cleavability and off-target effects as manifested by the increased-fidelity nucleases.
There is also a ranking among the targets by their cleavability and off-target effects manifested by the increased fidelity nucleases
Source:
usage constraintsupports
The three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers.
These three increased-fidelity nucleases can routinely be used only with perfectly matching 20 nucleotide-long spacers
Source:
Comparisons
Source-backed strengths
The available evidence places eSpCas9 within the class of increased-fidelity SpCas9 nucleases evaluated for specificity optimization. It was sufficiently well characterized to contribute mutations to HeFSpCas9 hybrid variants in comparative engineering studies.
Source:
No single nuclease variant shows generally superior fidelity
Source:
we generated new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
Source:
The paper concerns crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage.
Compared with LHCII N-terminal domain
eSpCas9 and LHCII N-terminal domain address a similar problem space because they share recombination.
Shared frame: same top-level item type; shared target processes: recombination; shared mechanisms: photocleavage
Strengths here: looks easier to implement in practice.
Compared with light-harvesting complex II
eSpCas9 and light-harvesting complex II address a similar problem space because they share recombination.
Shared frame: same top-level item type; shared target processes: recombination; shared mechanisms: photocleavage
Relative tradeoffs: appears more independently replicated.
Compared with SpCas9
eSpCas9 and SpCas9 address a similar problem space because they share recombination.
Shared frame: same top-level item type; shared target processes: recombination; shared mechanisms: dna binding, dna_binding, photocleavage
Relative tradeoffs: appears more independently replicated; looks easier to implement in practice.
Ranked Citations
- 1.