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

Cited 4 time in scopus
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Title
In vivo genome editing using the Cpf1 ortholog derived from Eubacterium eligens
Author(s)
Woo-Chan Ahn; Kwang Hyun Park; I S Bak; H N Song; Yan An; Su-Jin Lee; M Jung; K W Yoo; Dae Yeul Yu; Yong Sam Kim; B H Oh; Eui-Jeon Woo
Bibliographic Citation
Scientific Reports, vol. 9, pp. 13911-13911
Publication Year
2019
Abstract
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-γ gene, an essential gene for immunodeficient mouse model production, EeCpf1 efficiently generated IL2R-γ 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.
ISSN
2045-2322
Publisher
Springer-Nature Pub Group
DOI
http://dx.doi.org/10.1038/s41598-019-50423-6
Type
Article
Appears in Collections:
Critical Diseases Diagnostics Convergence Research Center > 1. Journal Articles
Synthetic Biology and Bioengineering Research Institute > Genome Editing Research Center > 1. Journal Articles
Division of Biomedical Research > Disease Target Structure Research Center > 1. Journal Articles
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