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double knock-in pig models with elements of binary Tet-On and phiC31 integrase systems for controllable and switchable gene expression

引用

Science China(Life Sciences) 2022年第11期65卷 2269-2286页

作者： 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 Kepin Wang CAS Key Laboratory of Regenerative Biology Joint School of Life SciencesGuangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesGuangzhou Medical UniversityGuangzhou 510530China Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine Centre for Regenerative Medicine and HealthHong Kong Institute of Science and InnovationGuangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesGuangzhou 510530China Sanya institute of Swine resource Guangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesHainan Provincial Research Center of Laboratory AnimalsSanya 572000China Research Unit of Generation of Large Animal disease Models Chinese Academy of Medical Sciences(2019RU015)Guangzhou 510530China University of Chinese Academy of Sciences Beijing 100049China Bioland Laboratory(Guangzhou Regenerative Medicine and Health Guangdong Laboratory) Guangzhou 510005China Guangdong Provincial Key Laboratory of Laboratory Animals Guangdong Laboratory Animals Monitoring InstituteGuangzhou 510633China Guangdong Provincial Key Laboratory of Large Animal models for Biomedicine Wuyi UniversityJiangmen 529020China

Inducible expression systems are indispensable for precise regulation and in-depth analysis of biological *** Tet-On system has been widely employed to regulate transgenic expression by *** pig models with tetracycline regulatory elements were generated through random *** process often resulted in uncertain expression and unpredictable phenotypes,thus hindering their ***,by precise knock-in of binary Tet-On 3G elements into Rosa26 and Hipp11 locus,respectively,a double knock-in reporter pig model was *** characterized excellent properties of this system for controllable transgenic expression both in vitro and in *** att P sites were arranged to flank the td Tomato to switch reporter *** or multiple gene replacement was efficiently and faithfully achieved in fetal fibroblasts and nuclear transfer *** display the flexible application of this system,we generated a pig strain with dox-inducing h KRASexpression through phiC31 integrase-mediated cassette *** eight months of dox administration,squamous cell carcinoma developed in the nose,mouth,and scrotum,which indicated this pig strain could serve as an ideal large animal model to study ***,the established pig models with controllable and switchable transgene expression system will provide a facilitating platform for transgenic and biomedical research.

关键词： double knock-in Tet-On system phiC31-mediated cassette exchange pig model hkrasg12d-related tumorigenesis

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