@article{8d747bab18e744a3a611b977dd8f4988,
title = "Chimeric CRISPR-CasX enzymes and guide RNAs for improved genome editing activity",
abstract = "A compact protein with a size of <1,000 amino acids, the CRISPR-associated protein CasX is a fundamentally distinct RNA-guided nuclease when compared to Cas9 and Cas12a. Although it can induce RNA-guided genome editing in mammalian cells, the activity of CasX is less robust than that of the widely used S. pyogenes Cas9. Here, we show that structural features of two CasX homologs and their guide RNAs affect the R-loop complex assembly and DNA cleavage activity. Cryo-EM-based structural engineering of either the CasX protein or the guide RNA produced two new CasX genome editors (DpbCasX-R3-v2 and PlmCasX-R1-v2) with significantly improved DNA manipulation efficacy. These results advance both the mechanistic understanding of CasX and its application as a genome-editing tool.",
keywords = "CRISPR, Cas12e, CasX, DNA cleavage, RNA-guided DNA nuclease, cryo-EM, genome editing, nucleic acid manipulation, sgRNA, structural engineering",
author = "Tsuchida, {Connor A.} and Shouyue Zhang and Doost, {Mohammad Saffari} and Yuqian Zhao and Jia Wang and Elizabeth O'Brien and Huan Fang and Li, {Cheng Ping} and Danyuan Li and Hai, {Zhuo Yan} and Jonathan Chuck and Julian Br{\"o}tzmann and Araz Vartoumian and David Burstein and Chen, {Xiao Wei} and Eva Nogales and Doudna, {Jennifer A.} and Liu, {Jun Jie Gogo}",
note = "Publisher Copyright: {\textcopyright} 2022 Elsevier Inc.",
year = "2022",
month = mar,
day = "17",
doi = "10.1016/j.molcel.2022.02.002",
language = "אנגלית",
volume = "82",
pages = "1199--1209.e6",
journal = "Molecular Cell",
issn = "1097-2765",
publisher = "Cell Press",
number = "6",
}