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Article

Identification of functional sgRNA mutants lacking canonical secondary structure using high-throughput FACS screening.

Cell reports - 25 Jun 2024

Liang Zeyu, Huang Chaoyong, Xia Yan, Ye Zhaojin, Fan Shunhua, Zeng Junwei, Guo Shuyuan, Ma Xiaoyan, Sun Lichao, Huo Yi-Xin

Abstract excerpt

Coexpressing multiple identical single guide RNAs (sgRNAs) in CRISPR-dependent engineering triggers genetic instability and phenotype loss. To provide sgRNA derivatives for efficient DNA digestion, we design a high-throughput digestion-activity-dependent positive screening strategy and astonishingly obtain functional nonrepetitive sgRNA mutants with up to 48 out of the 61 nucleotides mutated, and these...

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PMID: 38823012
DOI: 10.1016/j.celrep.2024.114290

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