Hypercompact adenine base editors based on a Cas12f variant guided by engineered RNA

Autor: Kim, Do Yon, Chung, Yuhee, Lee, Yujin, Jeong, Dongmin, Park, Kwang-Hyun, Chin, Hyun Jung, Lee, Jeong Mi, Park, Seyeon, Ko, Sumin, Ko, Jeong-Heon, Kim, Yong-Sam
Zdroj: Nature Chemical Biology; September 2022, Vol. 18 Issue: 9 p1005-1013, 9p
Abstrakt: Cas12f is a hypercompact type V, Cas12 family member. Previously, we reported a set of engineered guide RNAs supporting high indel efficiency for Cas12f1 in human cells. Here we suggest a new technology whereby the engineered guide RNAs also manifest high-efficiency programmable endonuclease activity for a Cas12f variant. We have termed this technology TaRGET (Tiny nuclease RNA-based Genome Editing Technology). Having this feature in mind, we established TaRGET-based adenine base editors (ABEs). A Tad–Tad mutant (V106W, D108Q) dimer fused to the C terminus of dCas12f (D354A) showed the highest levels of A-to-G conversion. The limited targetable sites for TaRGET-ABE were expanded with engineered variants of Cas12f or optimized deaminases. Delivery of TaRGET-ABE also ensured potent A-to-G conversion rates in mammalian genomes. Collectively, the TaRGET-ABE will contribute to improving precise genome-editing tools that can be delivered by adeno-associated viruses, thereby harnessing the development of clustered regularly interspaced short palindromic repeats (CRISPR)-based gene therapy.
Databáze: Supplemental Index
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