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Large Fragment Deletions Induced by Cas9 Cleavage While Not in BEs System in Rabbit

29 Pages Posted: 9 Aug 2019

See all articles by Yuning Song

Yuning Song

Jilin University (JLU) - Key Laboratory for Zoonosis Research; Jilin University (JLU) - Jilin Provincial Key Laboratory of Animal Embryo Engineering

Zhiquan Liu

Jilin University (JLU) - Jilin Provincial Key Laboratory of Animal Embryo Engineering

Yuxin Zhang

Jilin University (JLU) - Key Laboratory for Zoonosis Research; Jilin University (JLU) - Jilin Provincial Key Laboratory of Animal Embryo Engineering

Mao Chen

Jilin University (JLU) - Jilin Provincial Key Laboratory of Animal Embryo Engineering

Tingting Sui

Jilin University (JLU) - Jilin Provincial Key Laboratory of Animal Embryo Engineering

Liangxue Lai

Jilin University (JLU) - Key Laboratory for Zoonosis Research; Jilin University (JLU) - Jilin Provincial Key Laboratory of Animal Embryo Engineering

zhanjun li

Jilin University (JLU) - Jilin Provincial Key Laboratory of Animal Embryo Engineering

More...

Abstract

CRISPR-Cas9 and BEs system are poised to become the gene editing tool of choice in clinical contexts, however large fragment deletion was found in Cas9-mediated mutation cells without animal level validation. By analyzing 16 gene-edited rabbit lines (including 112 rabbits) generated using SpCas9, BEs, xCas9 and xCas9-BEs with long-range PCR genotyping and long-read sequencing by PacBio platform, we show that extending thousands of bases fragment deletions in single-guide RNA/Cas9 and xCas9 system mutation rabbit, but few large deletions were found in BEs-induced mutation rabbits. We firstly validated that no large fragment deletion induced by BEs system at animal level, suggesting that BE systems can be beneficial tools for the further development of highly accurate and secure gene therapy for the clinical treatment of human genetic disorders.

Funding Statement: This study was financially supported by the National Key Research and Development Program of China Stem Cell and Translational Research (2017YFA0105101). The Strategic Priority Research Program of the Chinese Academy of Sciences (XDA16030501, XDA16030503), Guangdong Province Science and Technology Plan Project (2014B020225003). China Postdoctoral Science Foundation Funded Project (2019T120236, 2018M641784).

Declaration of Interests: The authors declare that they have no competing interests.

Ethics Approval Statement: All animal studies were conducted according to the experimental practices and standards approved by the Animal Welfare and Research Ethics Committee at Jilin University.

Keywords: large fragment deletion, CRISPR/Cas9, BEs, Rabbits

Suggested Citation

Song, Yuning and Liu, Zhiquan and Zhang, Yuxin and Chen, Mao and Sui, Tingting and Lai, Liangxue and li, zhanjun, Large Fragment Deletions Induced by Cas9 Cleavage While Not in BEs System in Rabbit (August 7, 2019). Available at SSRN: https://ssrn.com/abstract=3433875 or http://dx.doi.org/10.2139/ssrn.3433875

Yuning Song

Jilin University (JLU) - Key Laboratory for Zoonosis Research

Changchun, 130062
China

Jilin University (JLU) - Jilin Provincial Key Laboratory of Animal Embryo Engineering

Jilin
China

Zhiquan Liu

Jilin University (JLU) - Jilin Provincial Key Laboratory of Animal Embryo Engineering

Changchun, 130062
China

Yuxin Zhang

Jilin University (JLU) - Key Laboratory for Zoonosis Research

Changchun, 130062
China

Jilin University (JLU) - Jilin Provincial Key Laboratory of Animal Embryo Engineering

Jilin
China

Mao Chen

Jilin University (JLU) - Jilin Provincial Key Laboratory of Animal Embryo Engineering

Jilin
China

Tingting Sui

Jilin University (JLU) - Jilin Provincial Key Laboratory of Animal Embryo Engineering

Jilin
China

Liangxue Lai

Jilin University (JLU) - Key Laboratory for Zoonosis Research ( email )

Changchun, 130062
China

Jilin University (JLU) - Jilin Provincial Key Laboratory of Animal Embryo Engineering

Changchun, 130062
China

Zhanjun Li (Contact Author)

Jilin University (JLU) - Jilin Provincial Key Laboratory of Animal Embryo Engineering ( email )

Changchun, 130062
China

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