Shanghai Biological Sciences in the discovery of the CRISPR/Cas12a cut ssDNA of the new activity

Source: Shanghai Biological Sciences in the discovery of the CRISPR/Cas12a cut ssDNA of new Active

3 month 12 days, the Chinese Academy of Sciences, molecular plant Sciences excellence and innovation center/Plant Physiology and ecology Research Institute Zhao screen of the study group of the latest research results, to CRISPR-Cas12a has both cis – and trans-cleavage activities on single-stranded DNA, the Cas12a both CIS-and TRANS-stranded DNA cleavage activity of as the title, published online in Cell Research. Papers in-depth systematic study of the Cas12a for the target single-stranded DNA and non-target single-stranded DNA cutting characteristics.

as life’s basic genetic material, DNA, the precise editing and rapid testing has been highly valued. In recent years, with the CRISPR/Cas9 system to discover and develop, awareness of gene therapy re-kindled a new hope; although there are some potential security risk, and a certain ethics of the dispute, the CRISPR/Cas9 system for which a relatively accurate and efficient, have begun to be applied to clinical research. At the same time, scientists began to the CRISPR system to introduce nucleic acids rapid detection in the field, it is desirable for the existing clinical diagnostic techniques bring about a breakthrough.

The study found Cas12a in the target ssDNA of the 22-bit near specific cleavage of DNA, called cis cut, and the cut behavior does not require the PAM sequence of the auxiliary. At the same time, the study also found that, once Cas12a with crRNA and target DNA to form a ternary complex, the complex will exhibit a strong“out-cut”activity, and the system in any single-stranded DNA cut into fragments are called trans cutting it. By means of mass spectrometry, the researchers found Cas12a the ssDNA and cut into 2-4nt fragments. According to a series of point mutations in the experimental results, the study proved Cas12a of the cis and trans cleavage activity and its RuvC pocket‘RuvC pocket’is relevant, and accordingly proposed Cas12a cut ssDNA molecules to the model. The research results are expected for DNA clinical diagnostics provide new research ideas.

In recent years, Zhao-screen Research Group is focused on synthetic biology enabling technology development and application, developed a number of DNA editing and detection methods, and to actively promote these methods for clinical research. Research work to obtain the CAS strategic pilot science and technology projects of“artificial chromosomes constructed with stable genetic technology method”, and the CAS youth innovation promotion Association, and the National Natural Science Fund Committee of the Fund.

CRIPSR/Cas12a cut single-stranded DNA a schematic diagram of A and a ternary complex structure of the simulation figure（B）。

Published at Fri, 15 Mar 2018 07:00:58 +0000