The CRISPR-Cas9 system has been adopted as a tool to manipulate the genomes of multiple living organisms, accelerating the pace of fundamental research and enabling clinical and agricultural breakthroughs. Besides, a newer type of genome editing approaches, named base-editor (BE) techniques, was recently developed. This kind of tool utilizes components from CRISPR systems together with other enzymes to directly install point mutations into cellular DNA or RNA. Nevertheless, base editors deriving from Cas9 nickase can cause unwanted DNA damage response (DDR).
Dr. Jia Chen’s team published the paper “Cas12a Base Editors Induce Efficient and Specific Editing with Low DNA Damage Response” in Cell Reports on June 2, 2020 (Wang Xiao, et al., 2020 ). In this article, the researchers demonstrated that the original version of catalytically dead Cas12a (dCas12a)-conjugated BEs induce a basal level of DNA breaks and minimally activate DDR proteins, including H2AX, ATM, ATR, and p53. They went on further to develop a dCas12a-based BEACON (base editing induced by human APOBEC3A and Cas12a without DNA break) system through fusing dCas12a with engineered human apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like 3A (APOBEC3A), and successfully applied it for both in vitro and in vivo editing. The BEACON system achieves efficient and specific base editing without generating DNA breaks or triggering DDR cascades, which is essential for its broad applications in mammalian cells and in clinics.
Plasmids from this article on MolecularCloud
Plasmids (pCmv-BEACON1 and pCmv-BEACON2) of this article have been deposited on MolecularCloud.
|MC_0101187||pCmv-BEACON1||Mammalian expression of BEACON1 human cells|
Mammalian expression of BEACON2 human cells
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