Targeted mutagenesis in mouse cells and embryos using an enhanced prime editor.
Soo-Ji ParkTae Yeong JeongSeung Kyun ShinDa Eun YoonSoo-Yeon LimSol Pin KimJungmin ChoiHyunji LeeJeong-Im HongJinhee AhnJe Kyung SeongKyoungmi KimPublished in: Genome biology (2021)
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.
Keyphrases
- crispr cas
- genome editing
- induced apoptosis
- gene expression
- dna damage
- mouse model
- cancer therapy
- stem cells
- genome wide
- signaling pathway
- metabolic syndrome
- cell therapy
- adipose tissue
- cell proliferation
- cell death
- insulin resistance
- binding protein
- skeletal muscle
- endoplasmic reticulum stress
- amino acid
- growth hormone