Source 1primary paper2017Genome biology
Toolkit/SpCas9 nucleases
SpCas9 nucleases
Also known as: enhanced SpCas9 nucleases, high fidelity SpCas9 nucleases
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
SpCas9 nucleases in this context are increased-fidelity CRISPR-Cas9 variants generated by combining mutations from eSpCas9 and SpCas9-HF1 into HeFSpCas9 forms. They are designed to preserve RNA-guided DNA cleavage while improving genome-editing specificity in a target-dependent manner.
Usefulness & Problems
Why this is useful
These nucleases are useful for applications requiring higher genome-editing specificity than standard SpCas9, particularly when off-target cleavage must be minimized. The source indicates that optimal use requires matching each target site 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
They address the engineering problem of balancing on-target cleavage activity with increased editing fidelity across different genomic targets. The cited work specifically tackles the lack of a single high-fidelity SpCas9 variant that is superior for all targets.
Source:
for highest specificity cleavage, each target needs to be matched with an appropriate high fidelity nuclease
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Component: A low-level protein part used inside a larger architecture that realizes a mechanism.
Mechanisms
mutation-mediated reduction of cleavage without equivalent reduction in dna bindingPhotocleavagerna-guided dna cleavageTechniques
No technique tags yet.
Target processes
No target processes tagged yet.
Implementation Constraints
Construct selection should account for target dependence, because the highest-specificity cleavage requires pairing each target with an appropriate high-fidelity nuclease. Guide design also matters: the source reports that a matching 5' G extension reduces activity more strongly than a mismatching 5' G extension. The provided evidence does not add further implementation details on expression system, delivery, or cofactor requirements.
No single high-fidelity SpCas9 nuclease variant is generally superior in fidelity across targets, which limits one-size-fits-all deployment. In addition, a matching 5' G extension is reported to be more detrimental to activity than a mismatching 5' G extension, indicating guide-format constraints for some targets.
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 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
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
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.
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
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 source1 linked approval claimfirst-pass slug spcas9-nucleases
Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage
Source:
optimization goalsupports
The paper concerns crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage.
Source:
Comparisons
Source-backed strengths
The tool class provides increased-fidelity SpCas9 variants assembled from eSpCas9 and SpCas9-HF1 mutations to optimize specificity and cleavage. The source further supports a practical design principle: different targets can benefit from different high-fidelity variants, enabling target-specific optimization.
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.
Ranked Citations
- 1.