Open AccessMultiplexed genome regulation in vivo with hyper-efficient Cas12a
Author(s) -
Lucie Y. Guo,
Jing Bian,
A. Edward Davis,
Pingting Liu,
H. Kempton,
Xiaowei Zhang,
Augustine Chemparathy,
Baokun Gu,
Xueqiu Lin,
Draven A. Rane,
Xiaoshu Xu,
Ryan M. Jamiolkowski,
Yang Hu,
Sui Wang,
Lei S. Qi
Publication year - 2022
Publication title -
nature cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 11.38
H-Index - 369
eISSN - 1476-4679
pISSN - 1465-7392
DOI - 10.1038/s41556-022-00870-7
Subject(s) - trans activating crrna , crispr , biology , genome engineering , genome editing , gene , microbiology and biotechnology , computational biology , cas9 , psychological repression , regulation of gene expression , gene expression , genetics
Multiplexed modulation of endogenous genes is crucial for sophisticated gene therapy and cell engineering. CRISPR-Cas12a systems enable versatile multiple-genomic-loci targeting by processing numerous CRISPR RNAs (crRNAs) from a single transcript; however, their low efficiency has hindered in vivo applications. Through structure-guided protein engineering, we developed a hyper-efficient Lachnospiraceae bacterium Cas12a variant, termed hyperCas12a, with its catalytically dead version hyperdCas12a showing significantly enhanced efficacy for gene activation, particularly at low concentrations of crRNA. We demonstrate that hyperdCas12a has comparable off-target effects compared with the wild-type system and exhibits enhanced activity for gene editing and repression. Delivery of the hyperdCas12a activator and a single crRNA array simultaneously activating the endogenous Oct4, Sox2 and Klf4 genes in the retina of post-natal mice alters the differentiation of retinal progenitor cells. The hyperCas12a system offers a versatile in vivo tool for a broad range of gene-modulation and gene-therapy applications.