1. Double knock-in pig models with elements of binary Tet-On and phiC31 integrase systems for controllable and switchable gene expression
- Author
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Qin, Jin, Xiaoyu, Yang, Shixue, Gou, Xiaoyi, Liu, Zhenpeng, Zhuang, Yanhui, Liang, Hui, Shi, Jiayuan, Huang, Han, Wu, Yu, Zhao, Zhen, Ouyang, Quanjun, Zhang, Zhaoming, Liu, Fangbing, Chen, Weikai, Ge, Jingke, Xie, Nan, Li, Chengdan, Lai, Xiaozhu, Zhao, Jiaowei, Wang, Meng, Lian, Lei, Li, Longquan, Quan, Yinghua, Ye, Liangxue, Lai, and Kepin, Wang
- Subjects
Male ,Animals, Genetically Modified ,Integrases ,Swine ,Animals ,Gene Expression ,Transgenes ,Genetic Therapy ,General Agricultural and Biological Sciences ,General Biochemistry, Genetics and Molecular Biology ,General Environmental Science - Abstract
Inducible expression systems are indispensable for precise regulation and in-depth analysis of biological process. Binary Tet-On system has been widely employed to regulate transgenic expression by doxycycline. Previous pig models with tetracycline regulatory elements were generated through random integration. This process often resulted in uncertain expression and unpredictable phenotypes, thus hindering their applications. Here, by precise knock-in of binary Tet-On 3G elements into Rosa26 and Hipp11 locus, respectively, a double knock-in reporter pig model was generated. We characterized excellent properties of this system for controllable transgenic expression both in vitro and in vivo. Two attP sites were arranged to flank the tdTomato to switch reporter gene. Single or multiple gene replacement was efficiently and faithfully achieved in fetal fibroblasts and nuclear transfer embryos. To display the flexible application of this system, we generated a pig strain with Dox-inducing hKRAS
- Published
- 2022