Novel Cas13 Proteins for Higher RNA Cleavage in CRISPR Editing
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Solution Overview
Problem
Existing CRISPR/Cas13 systems face challenges due to large molecular weight and low cleavage activity of Cas13 proteins, limiting their editing performance and RNA degradation efficiency, with a need for improved Cas13 proteins to enhance performance and functionality.
Innovation Solution
Development of novel Cas13 proteins, including Cas13a_10, Cas13a_11, Cas13b_3, Cas13b_4, Cas13b_5, Cas13T_9, Cas13T_15, Cas13T_16, and Cas13Z_2, with optimized amino acid sequences and potential fusion proteins, along with a CRISPR-Cas system incorporating guide RNAs for targeted RNA editing and detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If Cas13 proteins with large molecular weight are used, then RNA degradation capability is achieved, but cleavage activity is low and editing performance is impaired
Solution Approach 1:
The patent divides the Cas13 protein into smaller functional domains or uses truncated versions that retain essential RNA cleavage activity while removing bulky non-essential regions, thereby improving cellular uptake and editing performance without sacrificing RNA degradation capability
Solution Approach 2:
The patent modifies amino acid sequences of Cas13 proteins to optimize their physical and chemical properties, including reducing molecular weight while maintaining or enhancing catalytic activity through rational design and directed evolution approaches
2Reliability
If Cas13 proteins with large molecular weight are used, then RNA targeting function is maintained, but editing performance on target RNA is impaired
Solution Approach 1:
The patent extracts and removes non-essential bulky domains from Cas13 proteins while retaining the core RNA-guided cleavage machinery, improving cellular delivery and editing efficiency without compromising target recognition accuracy
Solution Approach 2:
The patent creates fusion proteins by combining Cas13 with other functional domains that enhance editing performance, such as nuclear localization signals or cell-penetrating peptides, to overcome the limitations of large molecular weight
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 novel Cas13 proteins exhibit enhanced RNA nuclease activity, facilitating efficient gene editing and highly sensitive nucleic acid detection, with applications in medical, biological, and environmental sample detection, and therapeutic uses for various diseases.
Implementation Method 1
the guide RNA is capable of binding to the Cas13 protein to form a complex and guiding the complex to contact a target RNA
Implementation Method 2
The novel Cas13 proteins exhibit enhanced RNA nuclease activity, facilitating efficient gene editing
Data Source
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AI summary
A Cas13 protein, a CRISPR-Cas system, and use thereof. The amino acid sequences of the Cas13 protein are as shown in SEQ ID NO. 1 to SEQ ID NO. 9, and are respectively named as Cas13a_10, Cas13a_11, Cas13b_3, Cas13b_4, Cas13b_5, Cas13T_9, Cas13T_15, Cas13T_16, and Cas13Z_2; these nine Cas13 proteins have low homology with the previously reported Cas13 proteins, and exhibit a RNA nuclease activity, showing great prospects in gene editing