CRISPR/Cas12J Enzyme With Simplified PAM Recognition for Gene Editing
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Solution Overview
Problem
Current CRISPR/Cas systems have limitations such as requiring multiple RNAs, varying sizes, complex PAM sequences, and potential off-target effects, necessitating the development of a more robust system for efficient gene editing.
Innovation Solution
A new CRISPR/Cas system featuring proteins with specific amino acid sequences (SEQ ID NOs: 1-20, 107, 108) and derivatives, capable of binding and cleaving target sequences under guide RNA guidance, with optional conjugation or fusion to enhance functionalities.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If Cas9, C2c1 or CasX is used for gene editing, then gene editing capability is achieved, but multiple guide RNAs are required which increases system complexity
Solution Approach 1:
The patent extracts and eliminates the requirement for multiple guide RNAs by designing a simplified CRISPR system that achieves gene editing capability with reduced RNA components, thereby reducing system complexity while maintaining adaptability
2Productivity
If Cpf1 is used for gene editing, then multiple gene editing is enabled with single guide RNA, but PAM sequence complexity increases
Solution Approach 1:
The patent changes the PAM sequence recognition parameters by designing new Cas proteins with simplified PAM requirements, allowing multiple gene editing operations while reducing the complexity of PAM sequence identification and selection
3Measurement precision
If C2c1 with 5'-TTN PAM recognition is used, then target site prediction becomes easier, but target site availability is reduced
Solution Approach 1:
The patent creates Cas proteins with universal PAM recognition capabilities that can identify multiple PAM sequence variants, thereby maintaining easy target site prediction while significantly expanding the availability of potential target sites across different genomes
4Productivity
If existing CRISPR/Cas systems are used, then gene editing function is achieved, but off-target effects occur which reduce precision
Solution Approach 1:
The patent incorporates feedback mechanisms through optimized Cas protein designs that enhance specificity recognition, allowing the system to distinguish more accurately between on-target and off-target sequences, thereby reducing off-target effects while maintaining gene editing functionality
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
The new system provides efficient and precise gene editing with reduced off-target effects, enabling improved nucleic acid editing capabilities.
Implementation Method 1
uses RNA guidance to specifically bind target sequences on the genome
Implementation Method 2
cut DNA to produce double-strand breaks
Data Source
AI summary
Provided are a Cas effector protein, a fusion protein containing said protein, and a nucleic acid molecule coding same. Also provided are a complex and a composition for nucleic acid editing, for example, a complex and a composition for gene or genome editing, containing the Cas effector protein or the fusion protein, or the nucleic acid molecule encoding same. Also provided is a method for nucleic acid editing, for example, a method for gene or genome editing, using the Cas effector protein or the fusion protein.