use of the 2a peptide for generation of multi-transgenic pigs through a single round of nuclear transfer使用2肽代multi-transgenic猪通过单个核移植.pdfVIP

Use of the 2A Peptide for Generation of Multi-Transgenic Pigs through a Single Round of Nuclear Transfer 1 1 1 1 1 1 1 Wei Deng , Dongshan Yang , Bentian Zhao , Zhen Ouyang , Jun Song , Nana Fan , Zhaoming Liu , Yu 1 2 2 1 3 2 1 Zhao , Qinghong Wu , Bayaer Nashun , Jiangjing Tang , Zhenfang Wu , Weiwang Gu *, Liangxue Lai * 1 Key Laboratory of Regenerative Biology, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China, 2 Institute of Comparative Medicine and Center of Laboratory Animals, Southern Medical University, Guangzhou, China, 3 College of Animal Science, South China Agricultural University, Guangzhou, China Abstract Multiple genetic modifications in pigs can essentially benefit research on agriculture, human disease and xenotransplan- tation. Most multi-transgenic pigs have been produced by complex and time-consuming breeding programs using multiple single-transgenic pigs. This study explored the feasibility of producing multi-transgenic pigs using the viral 2A peptide in the light of previous research indicating that it can be utilized for multi-gene transfer in gene therapy and somatic cell reprogramming. A 2A peptide-based double-promoter expression vector that mediated the expression of four fluorescent proteins was constructed and transfected into primary porcine fetal fibroblasts. Cell colonies (54.3%) formed under G418 selection co-expressed the four fluorescent proteins at uniformly high levels. The reconstructed embryos, which were