The cutting characteristics of Cas12a for target single-stranded DNA and non-target single-stranded DNA were systematically studied.

On March 12, the latest research from the Zhao Guoping Research Group of the Center for Excellence in Molecular Plant Sciences of the Chinese Academy of Sciences/Institute of Plant Physiology and Ecology was published online in Cell Research under the title CRISPR-Cas12a has both cis-and-trans-cleavage actives on-single-on-stranded DNA(Cas12a both conformive and trans-chain edited DNA activity).
the paper in-depth and systematic study of Cas12a for target single-stranded DNA and non-target single-stranded DNA cutting characteristics.
as the basic genetic material of life, DNA accurate editing and rapid detection has been highly valued.
in recent years, with the discovery and development of the CRISPR/Cas9 system, new hopes have been raised for gene therapy, and despite some potential safety risks and ethical battles, the CRISPR/Cas9 system has begun to be used in clinical research because of its relative accuracy and efficiency.
at the same time, scientists began to introduce the CRISPR system into the field of rapid detection of nucleic acids, hoping to bring about breakthroughs in existing clinical diagnostic techniques.
study found that Cas12a was specifically cut in the 22nd bit of the target ssDNA (called cis cutting), and that the cutting behavior did not require the aid of the PAM sequence.
, the study also found that once Cas12a and crRNA and target DNA form a ternary complex, the complex will show a strong "cut" activity, and any single-stranded DNA in the system is cut into fragments (called trans cutting).
using mass spectrometry, the researchers found cas12a cutting ssDNA into fragments of 2-4nt.
based on the experimental results of a series of point mutations, the study proves that cas12a's cis and trans cutting activity are related to its RuvC pocket ('RuvC pocket', and a molecular model of Cas12a cutting ssDNA is proposed.
the results of this research are expected to provide new research ideas for the clinical diagnosis of DNA.
in recent years, Zhao Guoping Research Group has focused on the development and application of synthetic biology enabling technologies, developed a number of DNA editing and testing methods, and actively promoted these methods for clinical research.
research has been supported by the Chinese Academy of Sciences' strategic leading science and technology project ("technical methods for artificial chromosome construction and stabilization of genetics"), the Chinese Academy of Sciences Youth Innovation Promotion Association and the National Natural Science Foundation.
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