Source 1review2018International Journal of Genomics
Toolkit/rational design
rational design
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Rational design is a protein engineering method used to improve or create novel protein functions. In the supplied evidence, it is discussed as one of the strategies used to expand CRISPR-Cas9 applications.
Usefulness & Problems
Why this is useful
This method is useful for engineering proteins when specific functional limitations constrain broader application. In the cited CRISPR-Cas9 context, the relevant limitations include off-target activity, strict protospacer-adjacent motif dependence, and the large size of Cas9 that complicates delivery.
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
Rational design helps address the need to modify protein properties that limit performance in biotechnology and disease-related applications. For CRISPR-Cas9 specifically, the evidence links these needs to reducing aberrant off-target activity, relaxing PAM constraints, and overcoming delivery problems associated with Cas9 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.
Published Workflows
Objective: Engineer AAV capsids for more precise and potent in vivo delivery to brain, muscle, and retina while reducing required vector dose.
Why it works: The abstract states that these approaches increasingly converge by integrating structural hypotheses, in vivo selections, and multi-trait computational optimization, implying that combining mechanistic design with empirical selection and computation improves capsid performance across tissues.
receptor-targeted BBB transport via TfR1 or ALPLfine-tuning HSPG interactions to facilitate ILM traversalliver detargeting for muscle-directed deliverydirected evolutionrational designmachine learningin vivo selectionsmulti-trait computational optimizationintegration of structural hypotheses
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete method used to build, optimize, or evolve an engineered system.
Mechanisms
No mechanism tags yet.
Target processes
No target processes tagged yet.
Implementation Constraints
The available evidence only states that rational design is used in studies that improve or develop novel protein functions. No practical details are provided on construct design, expression systems, delivery formats, cofactors, or experimental workflows.
The supplied evidence does not provide specific rationally designed constructs, quantitative performance data, or direct comparisons with other engineering methods. It also does not describe particular design rules, structural targets, or validation outcomes for any individual engineered protein.
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 rational-design
studies that improve or develop novel protein functions through ... rational design
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 rational design as a recognized strategy for improving or developing novel protein functions. It is positioned within protein engineering efforts aimed at expanding Cas9 utility across gene therapy, gene regulation, epigenome modification, and chromosome imaging.
Ranked Citations
- 1.