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Efficient deletion of LoxP-flanked selectable marker genes from the genome of transgenic pigs by an engineered Cre recombinase

文献类型： 外文期刊

作者： Huang, Xiaoling 1 ; Zou, Xian 1 ; Xu, Zhiqian 1 ; Tang, Fei 1 ; Shi, Junsong 4 ; Zheng, Engin 1 ; Liu, Dewu 1 ; Moisyadi, 1 ;

作者机构： 1.South China Agr Univ, Coll Anim Sci, Natl Engn Res Ctr Breeding Swine Ind, Guangzhou 510642, Guangdong, Peoples R China

2.South China Agr Univ, Coll Anim Sci, Guangdong Prov Key Lab Agroanim Genom & Mol Breed, Guangzhou 510642, Guangdong, Peoples R China

3.Guangdong Guangken Anim Husb Engn Res Inst Co Ltd, Guangzhou 510610, Guangdong, Peoples R China

4.Wens Foodstuff Grp Co Ltd, Guangdong Wens Pig Breeding Technol Co Ltd, Yunfu 527400, Guangdong, Peoples R China

5.Univ Hawaii Manoa, John A Burns Sch Med, Inst Biogenesis Res, Honolulu, HI 96822 USA

6.Manoa BioSci, 1717 Mott Smith Dr 3213, Honolulu, HI 96822 USA

关键词： Selectable marker-free; pigs; Cre; LoxP; HTNCre; SCNT

期刊名称：TRANSGENIC RESEARCH （ 影响因子：2.788； 五年影响因子：2.377 ）

ISSN： 0962-8819

年卷期： 2020 年 29 卷 3 期

页码：

收录情况： SCI

摘要： Genetically modified (GM) pigs hold great promises for pig genetic improvement, human health and life science. When GM pigs are produced, selectable marker genes (SMGs) are usually introduced into their genomes for host cell or animal recognition. However, the SMGs that remain in GM pigs might have multiple side effects. To avoid the possible side effects caused by the SMGs, they should be removed from the genome of GM pigs before their commercialization. The Cre recombinase is commonly used to delete the LoxP sites-flanked SMGs from the genome of GM animals. Although SMG-free GM pigs have been generated by Cre-mediated recombination, more efficient and cost-effective approaches are essential for the commercialization of SMG-free GM pigs. In this article we describe the production of a recombinant Cre protein containing a cell-penetrating and a nuclear localization signal peptide in one construct. This engineered Cre enzyme can efficiently excise the LoxP-flanked SMGs in cultured fibroblasts isolated from a transgenic pig, which then can be used as nuclear donor cells to generate live SMG-free GM pigs harboring a desired transgene by somatic cell nuclear transfer. This study describes an efficient and far-less costly method for production of SMG-free GM pigs.

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