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Targeted mutagenesis in mouse cells and embryos using an enhanced prime editor

Cited 13 time in Web of Science Cited 14 time in Scopus
Issue Date
2021-06-03
Publisher
BMC
Citation
Genome Biology. 2021 Jun 03;22(1):170
Keywords
Prime editorIgf2Adamts20Mouse cells and embryosGermline transmissionDwarf phenotypeProximal dead sgRNAChromatin-modulating peptides
Abstract
Prime editors, novel genome-editing tools consisting of a CRISPR-Cas9 nickase and an engineered reverse transcriptase, can induce targeted mutagenesis. Nevertheless, much effort is required to optimize and improve the efficiency of prime-editing. Herein, we introduce two strategies to improve the editing efficiency using proximal dead sgRNA and chromatin-modulating peptides. We used enhanced prime-editing to generate Igf2 mutant mice with editing frequencies of up to 47% and observed germline transmission, no off-target effects, and a dwarf phenotype. This improved prime-editing method can be efficiently applied to cell research and to generate mouse models.
ISSN
1474-760X
Language
English
URI
https://hdl.handle.net/10371/174784
DOI
https://doi.org/10.1186/s13059-021-02389-w
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College of Veterinary Medicine (수의과대학)Dept. of Veterinary Medicine (수의학과)Journal Papers (저널논문_수의학과)
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