Open AccessEfficient deletion of LoxP-flanked selectable marker genes from the genome of transgenic pigs by an engineered Cre recombinase
Author(s) -
Xiaoling Huang,
Xian Zou,
Zhiqian Xu,
Fei Tang,
Junsong Shi,
Enqin Zheng,
Dewu Liu,
Stefan Moisyadi,
Johann Urschitz,
Zhenfang Wu,
Zicong Li
Publication year - 2020
Publication title -
transgenic research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.724
H-Index - 83
eISSN - 1573-9368
pISSN - 0962-8819
DOI - 10.1007/s11248-020-00200-3
Subject(s) - cre recombinase , biology , cre lox recombination , selectable marker , transgene , recombinase , genome , genetically modified organism , gene , transgenesis , genetics , gene targeting , microbiology and biotechnology , computational biology , genetically modified mouse , recombination , reproductive biology , embryogenesis
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.