Journal
GENOME BIOLOGY
Volume 22, Issue 1, Pages -Publisher
BMC
DOI: 10.1186/s13059-021-02389-w
Keywords
Prime editor; Igf2; Adamts20; Mouse cells and embryos; Germline transmission; Dwarf phenotype; Proximal dead sgRNA; Chromatin-modulating peptides
Funding
- Chung Yang, Cha Young Sun, AMP
- Jang Hi Joo Memorial Fund
- Bio AMP
- Medical Technology Development Program of the National Research Foundation (NRF) of Korea (Korea Mouse Phenotyping Project) [NRF-2013M3A9D5072550, NRF-2020M3A9D5A01082439, NRF2019R1A2C2087198, NRF-2019M3A9H1103792]
- National Research Foundation of Korea [2020M3A9D5A01082439, 2019M3A9H1103792] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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Prime editors, a novel genome-editing tool, can induce targeted mutations with high efficiency. By utilizing two strategies to improve editing efficiency, we successfully generated Igf2 mutant mice with editing frequencies of up to 47% and observed important phenotypic changes.
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.
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