Source 1primary paper2017Genome biology
Toolkit/HeFSpCas9
HeFSpCas9
Also known as: HeFSpCas9s, Highly enhanced Fidelity nuclease variants
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
HeFSpCas9 denotes engineered Streptococcus pyogenes Cas9 high-fidelity nuclease variants that combine mutations from eSpCas9 and SpCas9-HF1. These variants were developed to optimize the balance between on-target cleavage activity and genome-editing specificity across different target sites.
Usefulness & Problems
Why this is useful
HeFSpCas9 is useful as a panel of increased-fidelity SpCas9 nucleases for applications where off-target cleavage must be reduced while retaining sufficient target cleavage. The source indicates that highest-specificity editing is achieved by matching each genomic target with an appropriate high-fidelity nuclease variant rather than relying on a single universally best enzyme.
Source:
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
Problem solved
This tool addresses the engineering problem that no single high-fidelity SpCas9 variant is generally superior in fidelity across all targets. It therefore helps solve the need to tune the activity-specificity tradeoff for individual guide RNA and target combinations.
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
DerivedHeFSpCas9 denotes engineered Streptococcus pyogenes Cas9 high-fidelity nuclease variants created by combining mutations from eSpCas9 and SpCas9-HF1. These variants were developed to optimize the balance between target cleavage activity and specificity across genomic targets.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Component: A low-level protein part used inside a larger architecture that realizes a mechanism.
Mechanisms
DNA Bindingdna cleavagedna cleavagePhotocleavagerna-guided dna bindingrna-guided dna bindingTechniques
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
HeFSpCas9 variants are engineered SpCas9 proteins containing combined mutations from eSpCas9 and SpCas9-HF1. Practical use requires empirical target-by-target selection of the most appropriate high-fidelity nuclease, and guide designs with 5' G extensions should be considered carefully because matching 5' G extensions can reduce activity.
The evidence indicates that no single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets, so HeFSpCas9 performance is target dependent. The source also reports that a matching 5' G extension is more detrimental to activity than a mismatching 5' G extension for increased-fidelity nucleases, which constrains guide design.
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 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 source9 linked approval claimsfirst-pass slug hefspcas9
new "Highly enhanced Fidelity" nuclease variants (HeFSpCas9s) containing mutations from both eSpCas9 and SpCas9-HF1
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:
method frameworksupports
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
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
A key strength is that HeFSpCas9 variants explicitly combine mutations from two established high-fidelity SpCas9 designs, eSpCas9 and SpCas9-HF1. The cited study concluded that target-dependent matching of high-fidelity nucleases can provide the highest-specificity cleavage, supporting the value of this crossed-variant strategy.
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 eSpCas9
HeFSpCas9 and eSpCas9 address a similar problem space because they share recombination.
Shared frame: same top-level item type; shared target processes: recombination; shared mechanisms: dna cleavage, dna_binding, photocleavage
Compared with LHCII N-terminal domain
HeFSpCas9 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 SpCas9
HeFSpCas9 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, photocleavage
Relative tradeoffs: appears more independently replicated; looks easier to implement in practice.
Ranked Citations
- 1.