Source 1review2018International Journal of Genomics
Toolkit/protein splitting
protein splitting
Also known as: domain splitting
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Protein splitting is a protein engineering method in which proteins are modified through domain fusion or splitting to improve existing functions or develop novel functions. In the provided evidence, it is discussed as a strategy relevant to expanding CRISPR-Cas9 applications.
Usefulness & Problems
Why this is useful
This method is useful as a general protein engineering approach for modifying protein function. In the cited CRISPR-Cas9 context, it is relevant because Cas9 is used in gene therapy, gene regulation, epigenome modification, and chromosome imaging, yet its broader use is constrained by off-target activity, PAM dependence, and delivery challenges associated with large protein size.
Source:
CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as gene therapy, gene regulation, modifying epigenomes, and imaging chromosomes.
Problem solved
In the provided evidence context, protein splitting is positioned as an engineering strategy for addressing limitations that restrict CRISPR-Cas9 deployment. Specifically, the motivating problems named are aberrant off-target activity, strict protospacer-adjacent motif dependence, and delivery problems caused by the large size of Cas9.
Source:
CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as gene therapy, gene regulation, modifying epigenomes, and imaging chromosomes.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete method used to build, optimize, or evolve an engineered system.
Target processes
No target processes tagged yet.
Implementation Constraints
The only implementation detail supported by the evidence is that the method operates through domain fusion or domain splitting. No construct architecture, linker design, host organism, delivery modality, or assay conditions are provided in the supplied material.
The supplied evidence does not describe a specific split-protein construct, experimental system, or validated outcome for this method. It also does not provide direct evidence that protein splitting alone resolves Cas9 off-targeting, PAM constraints, or delivery barriers in a demonstrated implementation.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
CRISPR-Cas9 has been used across applications including gene therapy, gene regulation, epigenome modification, and chromosome imaging.
CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as gene therapy, gene regulation, modifying epigenomes, and imaging chromosomes.
These Cas9 limitations hinder the use of CRISPR for disease treatment and wider biotechnological applications.
These obstacles hinder the use of CRISPR for disease treatment and in wider biotechnological applications.
Cas9 can exhibit aberrant off-target activity.
(ii) aberrant off-target activity
Cas9 has a strict dependence on a protospacer-adjacent motif sequence.
some limitations have also been reported, for instance (i) a strict dependence on a protospacer-adjacent motif (PAM) sequence
Cas9 lacks sufficient modulation of protein binding and endonuclease activity for precise spatiotemporal control.
(iv) lack of modulation of protein binding and endonuclease activity, which is crucial for precise spatiotemporal control of gene expression or genome editing
The large size of Cas9 creates problems for CRISPR delivery.
(iii) the large size of Cas9 is problematic for CRISPR delivery
The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.
Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.
Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.
Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.
Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.
Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.
Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.
Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.
Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.
Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.
Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.
Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.
Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.
Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.
Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Approval Evidence
1 source2 linked approval claimsfirst-pass slug protein-splitting
studies that improve or develop novel protein functions through domain fusion or splitting
Source:
review focussupports
The review emphasizes domain fusion or splitting, rational design, and directed evolution as protein-engineering strategies for expanding SpCas9 versatility.
Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
Source:
strategy overviewsupports
Protein-engineering approaches are presented as solutions to overcome Cas9 limitations and generate more robust and efficient DNA manipulation tools.
Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Source:
Comparisons
Source-backed strengths
The evidence supports that protein splitting can be used to improve protein function or generate novel protein functions through domain-level reconfiguration. It is also presented within a framework of strategies intended to expand CRISPR-Cas9 applications, but no quantitative performance data or direct comparative results are provided here.
Ranked Citations
- 1.