一种融合RNA结合结构域的工程化的Cas13d蛋白及其应用
By inserting an RNA-binding domain into the Cas13d protein and incorporating mismatch design, the problem of balancing specificity and sensitivity in single nucleotide variant detection in CRISPR was solved, achieving efficient, low-dependency, and widely applicable SNV detection.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
- Filing Date
- 2026-03-25
- Publication Date
- 2026-07-17
AI Technical Summary
Existing CRISPR nucleic acid detection technology faces technical bottlenecks in identifying single nucleotide variants (SNVs), including a balance between specificity and sensitivity, sluggish reaction kinetics, high amplification dependence, and limited site designability.
Develop an engineered Cas13d protein fused with an RNA-binding domain (RBD). By inserting a heterologous RNA-binding domain into the Cas13d protein at a specific site, the efficiency of target RNA binding and activation can be improved. Combined with mismatch design to enhance single-base discrimination ability, and optionally equipped with a reverse transcription loop-mediated isothermal amplification (RT-LAMP) module to reduce amplification dependence.
This improved the sensitivity and stability of the CRISPR system in single-base discrimination, reduced dependence on amplification, expanded the feasibility of site design, simplified the detection process, and improved the reliability and sensitivity of detection.
Smart Images
Figure CN121896201B_ABST