Continuous Cas13 nuclease activity may cause unpredictable off-target, hindering the popularization and application of gene editing technology CRISPR. Researchers from the Army Medical Center of the Army Medical University and other units have discovered that the inhibitor of Cas13a nuclease activity, anti-CRISPR (AcrVIAs), can turn off the CRISPR-Cas13a RNA editing system. The research solved the problems that hindered the popularization and application of CRISPR. Related results have been published online in the journal Molecular Cells recently.
As an emerging RNA editing technology, CRISPR-Cas13 can achieve target gene knockdown and base-point editing without changing the genome, and analyze the function of genes , Disease diagnosis, human disease targeted therapy and other fields have great potential and broad application prospects. However, the existing CRISPR is not perfect. “One is that gene editing cannot freely control the time and intensity, and more importantly, the continuous Cas13 nuclease activity may cause unpredictable off-target effects, which restricts the promotion and application of this technology.” Army Military Medical University Army Special Medical Center War Injury Jiang Jianxin, director of the research frontier technology research department, said.
By establishing the theory and method of bioinformation analysis for AcrVIAs identification, researchers quickly discovered multiple AcrVIAs that inhibit Cas13a nuclease. After in vitro transcription and translation experiments, as well as phage plaque killing experiments, the study confirmed that the AcrVIAs protein helped the phage successfully escape the CRISPR-Cas13a system.
“Our comprehensive research on RNA editing inhibition in mammals and human cells has confirmed that these AcrVIAs can be used as effective inhibitors of RNA editing in human cells, This will reduce the off-target effect caused by Cas13 nuclease activity and improve the accuracy of gene editing. “Said Dr. Lin Ping, the first author of the paper and the frontier technology research laboratory of the Army Medical Center of the Army Military Medical University.
The newly discovered anti-Cas13a provides a switch for controlling Cas13a in gene expression regulation, disease diagnosis and treatment. Once gene editing is completed, Cas13a activity can be eliminated immediately . Jiang Jianxin introduced that this discovery will make the clinical application of Cas13a RNA editor more extensive, more precise and less side effects.
(Original title My scientist discovered a new inhibitor that can improve the accuracy of gene editing)