Identification of ACTB Gene as a Potential Safe Harbor Locus in Pig Genome
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ORIGINAL PAPER
Identification of ACTB Gene as a Potential Safe Harbor Locus in Pig Genome Youcai Xiong1 · Xiaosong Han1 · Jinfu Zhang1 · Guangxing Zhao1 · Zichang Wang1 · Rongzhi Zhuang1 · Xiongwei Nie1 · Shengsong Xie1,2 · Changchun Li1,2 · Xinyun Li1,2 · Xiangdong Liu1,2 · Kui Li3 · Shuhong Zhao1,2 · Jinxue Ruan1,4 Accepted: 18 September 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Transgenic pigs play an important role in biomedicine and agriculture. The “safe harbor” locus maintains consistent foreign gene expression in cells and is important for transgenic pig generation. However, as only several safe harbor loci(Rosa26, pH11 and Pifs501) have been identified in pigs, meeting the needs of the insertion of various foreign genes is difficult. In this study, we develop a novel strategy for the efficient knock-in of gene-of-interest fragments into endogenous beta-actin(ACTB) gene via CRISPR/Cas9 mediated homologous recombination with normal expression of ACTB. Thus, we provide an alternative strategy to integrate exogenous genes into the pig genome that can be applied to agricultural breeding and biomedical models. Keywords Safe harbor · ACTB · CRISPR/Cas9 · Homologous recombination · Pig
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12033-020-00276-6) contains supplementary material, which is available to authorized users. * Jinxue Ruan [email protected]
Xinyun Li [email protected]
Youcai Xiong [email protected]
Xiangdong Liu [email protected]
Xiaosong Han [email protected]
Kui Li [email protected]
Jinfu Zhang [email protected]
Shuhong Zhao [email protected]
Guangxing Zhao [email protected]
1
Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Huazhong Agricultural University, Ministry of Education, Wuhan 430070, China
2
The Cooperative Innovation Center for Sustainable Pig Production‑Swine Breeding and Reproduction Innovation Platform, Huazhong Agricultural University, Wuhan 430070, China
3
State Key Laboratory of Animal Nutrition, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China
4
College of Life Science, Foshan University, Foshan 528231, Guangdong, China
Zichang Wang [email protected] Rongzhi Zhuang [email protected] Xiongwei Nie [email protected] Shengsong Xie [email protected] Changchun Li [email protected]
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Introduction Transgenic pigs play an important role in improving human health when used as animal models for various human diseases in biomedicine and production of highquality food in agriculture [1–4]. The expression level of the foreign genes is important in the applications of those transgenic products [5]. The “safe harbor” locus maintains the consistent expression of foreign gene expressing in cells. To date, however, only several safe harbor loci, namely, pRosa26, pH11 and Pifs501, have been identified in pig
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