CRISPR-Cas12a: Opens the door to efficient gene editing for you Genome Biology。

Title: Enhanced into the journal: Enhanced into the world by new Cas12a orthologs with optimized crRNA scaffolds
Journal:
Fei Teng†, Jing Li†, Tong Cuitong†, Kai Xu, Lu Guo, Qingqin Gao, Guihai Feng, Chuanyuan Chen, Dali Han, Qi Zhou and Wei Li
Published: 2019/02/05
Digital Identification Number: 10.1186/s13059-019-1620-8
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Wei Li is a researcher at the Institute of Zoology of the Chinese Academy of Sciences. His team focuses on the field of reproduction and regeneration, as well as the development of genomic engineering tools and the exploration of new approaches to gene therapy; Fei Teng is a Ph.D. candidate at the Institute of Zoology of the Chinese Academy of Sciences. His current research focuses on the development and application of CRISPR-Cas systems for disease modeling and new genome editing tools.
recently published in
, the researchers found several new CRISPR-Cas12a gene constellations, a CRISPR-Cas system used to edit mammalian genomes. The two authors, Wei Li and Fei Teng, showed us their research and how the findings enhance and expand gene editing tools.in just a few years, a technique called CRISPR has changed not only the way scientists make specific changes to DNA, but also the way people might think about treating diseases. At the same time, the emergence of CRISPR-Cas systems in the microbial world has accelerated the emergence of new gene editing tools.
The study, published in
by Dr. Wei Li's team from the Institute of Zoology of the Chinese Academy of Sciences, provides us with a more practical Cas12a/Cpf1 cogeneration, expands our selection of CRISPR-Cas-based systems for gene editing, and enhances the targeting efficiency of Cas12a enzymes through genetic engineering methods and optimizing the casffold structure of CRISPR RNA (crRNA).
technology mediated by CRISPR-Cas is a promising field with broad applications in basic research and biomedical applications. To date, six types and more than 20 CRISPR-Cas system subtypes have been found. CRISPR-Cas12a is a class II (V-type) CRISPR-Cas system used to edit mammalian genomes. Cas12a has unique features that complement crispr-Cas9 systems. However, the system still has some drawbacks that need to be improved, such as lower genomic coverage and lower targeting efficiency.
the study, Wei Li and his team dug deep into the database and identified several new CRISPR-Cas12a gene constellations that have been shown to be used for gene editing in mammalian cells. In this article, they delved into the editing of these six new Cas12a co-origins.。 Wei Li et al. found that the V-A Cas12a system is very conservative, especially in mature crRNA sequences and secondary structures. Based on this conservatism, the researchers concluded that the PAM sequences they needed were also conservative. As the in-body study data in this paper reveal, these newly identified Cas12a nucleases were able to utilize the 5T-PAM rich region, and the authors then discovered more potential site for Cas12a nucleases to edit mammalian genomes.compared to the widely used AsCas12a and LbCas12a, some of the six Cas12a cogeners can identify simpler 5-TTN PAMs, raising genome coverage to four times. In addition, in in-body cutting tests, they found that HkCas12a could utilize unconventional PAMs (5-YYN and 5-TBBN-). Although few PAM sequences have been identified in cell line, researchers can use HkCas12a to identify 5'-YTN and 5'-TYYN PAMs for gene editing in mammals. In the mammalian genome, hkCas12a's recognition of PAM can triple hkCas12a's target range compared to the classic 5-TTN PAM.study, Wei Li and his team observed that different crRNA scaffolds can affect the targeting efficiency of the Cas12a protein. In most cases, the activity of cas12a protein is reduced or even eliminated completely, but in rare cases, some changes in the crRNA scaffolds ring region can increase the activity of Cas12a nuclease. They successfully screened a variant (crRNA4n96) that significantly improved the efficiency of gene editing mediated by Cas12a.
, the study found six new Cas12a that could be used for gene editing, with higher genomic coverage and more gene editing options for researchers. More importantly, by designing and optimizing crRNA scaffolds, the targeting efficiency of Cas12a isoens can be effectively improved.。 CRISPR-Cas12a/Cpf1, a single RNA-guided endonuclease system, provides a promising tool for genome engineering. However, only three Cas12a orthologs have been employed for mammalian genome editing, and the editing efficiency as well as targeting coverage still requires improvements. Here, we harness six novel Cas12a orthologs for genome editing in human and mouse cells, some of which utilize simple protospacer adjacent motifs (PAMs) that remarkably increase the targeting range in the genomes. Moreover, we identify optimized CRISPR RNA (crRNA) scaffolds that can increase the genome editing efficiency of Cas12a.。 ( ) publishes outstanding research in all areas of biology and biomedicine studied from a genomic and post-genomic perspective.
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(Source: Science.com)