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Targeted mutagenesis in mice via an engineered AsCas12f1 system.

ABSTRACT: SpCas9 and AsCas12a are widely utilized as genome editing tools in human cells, but their applications are largely limited by their bulky size. Recently, AsCas12f1 protein, with a small size (422 amino acids), has been demonstrated to be capable of cleaving double-stranded DNA protospacer adjacent motif (PAM). However, low editing efficiency and large differences in activity against different genomic loci have been a limitation in its application. Here, we show that engineered AsCas12f1 sgRNA has significantly improved the editing efficiency in human cells and mouse embryos. Moreover, we successfully generated three stable mouse mutant disease models using the engineered CRISPR-AsCas12f1 system in this study. Collectively, our work uncovers the engineered AsCas12f1 system expands mini CRISPR toolbox, providing a remarkable promise for therapeutic applications.

SUBMITTER: Fan P

PROVIDER: S-EPMC10821844 | biostudies-literature | 2024 Jan

REPOSITORIES: biostudies-literature

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Targeted mutagenesis in mice via an engineered AsCas12f1 system.

Fan Peng P Wang Hejun H Zhao Feiyu F Zhang Tao T Li Jinze J Sun Xiaodi X Yu Yongduo Y Xiong Haoyang H Lai Liangxue L Sui Tingting T

Cellular and molecular life sciences : CMLS 20240128 1

SpCas9 and AsCas12a are widely utilized as genome editing tools in human cells, but their applications are largely limited by their bulky size. Recently, AsCas12f1 protein, with a small size (422 amino acids), has been demonstrated to be capable of cleaving double-stranded DNA protospacer adjacent motif (PAM). However, low editing efficiency and large differences in activity against different genomic loci have been a limitation in its application. Here, we show that engineered AsCas12f1 sgRNA ha ...[more]

PMID: 38280977

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