학술논문

Targeted mutagenesis in mouse cells and embryos using an enhanced prime editor
Document Type
article
Source
Genome Biology, Vol 22, Iss 1, Pp 1-11 (2021)
Subject
Prime editor
Igf2
Adamts20
Mouse cells and embryos
Germline transmission
Dwarf phenotype
Biology (General)
QH301-705.5
Genetics
QH426-470
Language
English
ISSN
1474-760X
Abstract
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.