In vivo genome editing using the Cpf1 ortholog derived from Eubacterium eligens

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Cpf1 is an RNA-guided endonuclease that can be programmed to cleave DNA targets. Specific features, such as containing a short crRNA, creating a staggered cleavage pattern and having a low off-target rate, render Cpf1 a promising gene-editing tool. Here, we present a new Cpf1 ortholog, EeCpf1, as a genome-editing tool; this ortholog is derived from the gut bacterial species Eubacterium eligens. EeCpf1 exhibits a higher cleavage activity with the Mn2+ metal cofactor and efficiently cuts the target DNA with an engineered, nucleotide extended crRNA at the 5' target site. When mouse blastocysts were injected with multitargeting crRNAs against the IL2R-gamma gene, an essential gene for immunodeficient mouse model production, EeCpf1 efficiently generated IL2R-gamma knockout mice. For the first time, these results demonstrate that EeCpf1 can be used as an in vivo gene-editing tool for the production of knockout mice. The utilization of engineered crRNA with multiple target sites will help to explore the in vivo DNA cleavage activities of Cpf1 orthologs from other species that have not been demonstrated.
Publisher
NATURE PUBLISHING GROUP
Issue Date
2019-09
Language
English
Article Type
Article
Citation

SCIENTIFIC REPORTS, v.9

ISSN
2045-2322
DOI
10.1038/s41598-019-50423-6
URI
http://hdl.handle.net/10203/269971
Appears in Collection
BS-Journal Papers(저널논문)
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