Cas-phi Variant Polypeptides Enhance Nuclease Activity
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Solution Overview
Problem
Current CRISPR/Cas systems face challenges in achieving efficient in vitro detection and effective in vivo genome engineering, necessitating the exploration of alternative strategies and components.
Innovation Solution
The development of variant polypeptides and guide nucleic acids that leverage nucleic acid modifying activities for the modification and detection of target nucleic acids, including engineered polypeptides with enhanced nuclease activity and binding affinity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If native CRISPR/Cas systems are used, then sequence-specific targeting capability is achieved, but nuclease activity and binding affinity are insufficient for efficient in vitro detection and in vivo genome engineering
Solution Approach 1:
The patent applies parameter changes by systematically mutating specific amino acid residues in the Cas protein sequence to optimize its biochemical properties. Through high-throughput screening of variant libraries, the invention identifies mutations that enhance nuclease activity, binding affinity, and overall performance for both in vitro detection and in vivo genome engineering applications.
2Reliability
If native CRISPR/Cas systems are used, then genome editing capability is achieved, but binding affinity and specificity are insufficient for effective in vivo applications
Solution Approach 1:
The patent employs parameter changes by mutating amino acid residues involved in guide RNA binding and target recognition. The engineered variants demonstrate improved binding affinity and specificity, enabling more effective and efficient in vivo genome engineering applications.
3Productivity
If conventional polypeptide sequences are used, then basic CRISPR function is maintained, but enhanced performance for detection and therapy is not achieved
Solution Approach 1:
The patent applies parameter changes by optimizing specific amino acid positions while maintaining overall protein structure and function. This approach enhances detection sensitivity and therapeutic performance without requiring complete redesign of the polypeptide sequence.
Solution Approach 2:
The patent uses copying by creating variant libraries based on the native Cas protein sequence. These variants are screened to identify copies with improved properties, allowing propagation of beneficial mutations while maintaining the core functional architecture.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
These variant polypeptides and guide nucleic acids enable precise modification and detection of target nucleic acids, potentially leading to improved genome editing and therapeutic applications.
Implementation Method 1
CRISPR/Cas systems provide immunity in bacteria and archaea against viruses and plasmids by targeting the nucleic acids of the viruses and plasmids in a sequence-specific manner
Implementation Method 2
Compositions, systems, and methods disclosed herein leverage nucleic acid modifying activities (e.g., cis cleavage activity) of these polypeptides and guide nucleic acids for the modification and detection of target nucleic acids
Data Source
AI summary
Provided herein are compositions, systems, and methods comprising engineered effector proteins and uses thereof. These effector proteins may be characterized as CRISPR-associated (Cas) proteins. Various compositions, systems, and methods of the present disclosure may leverage the activities of these effector proteins for the modification, detection, and engineering of nucleic acids.