Use of the 2A peptide for generation of multi-transgenic pigs through a single round of nuclear transfer.
Use of the 2A peptide for generation of multi-transgenic pigs through a single round of nuclear transfer.
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DOI：
10.1371/journal.pone.0019986
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Lai L
中科院分区：
文献类型：
--
作者：
Deng W;Yang D;Zhao B;Ouyang Z;Song J;Fan N;Liu Z;Zhao Y;Wu Q;Nashun B;Tang J;Wu Z;Gu W;Lai L
猪的多种遗传修饰可以从根本上促进农业、人类疾病和异种移植的研究。大多数多转基因猪都是通过使用多个单转基因猪的复杂且耗时的育种程序产生的。本研究探讨了利用病毒2A肽生产多转基因猪的可行性，并指出其可用于基因治疗和体细胞重编程中的多基因转移。构建了介导4种荧光蛋白表达的2A肽双启动子表达载体，并转染原代猪胎儿成纤维细胞。在G418选择下形成的细胞集落（54.3%）以均匀的高水平共表达四种荧光蛋白。将通过体细胞核移植获得并确认均匀表达四种荧光蛋白的重构胚胎移植到七头受体后备母猪体内。11头仔猪由2头后备母猪分娩，其中7头在各种组织中以同等高水平共表达4种荧光蛋白。使用护目镜直接观察鼻、蹄和舌处的荧光强度。结果表明，2A肽和双启动子相结合的策略有效地介导了四种荧光蛋白在猪中的共表达，因此是一种有前途的方法，通过一次核转移产生多个转基因猪。
Multiple genetic modifications in pigs can essentially benefit research on agriculture, human disease and xenotransplantation. 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 obtained by somatic cell nuclear transfer and confirmed to express the four fluorescent proteins evenly, were transplanted into seven recipient gilts. Eleven piglets were delivered by two gilts, and seven of them co-expressed the four fluorescent proteins at equivalently high levels in various tissues. The fluorescence intensities were directly observed at the nose, hoof and tongue using goggles. The results suggest that the strategy of combining the 2A peptide and double promoters efficiently mediates the co-expression of the four fluorescent proteins in pigs and is hence a promising methodology to generate multi-transgenic pigs by a single nuclear transfer.
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影响因子：
2.2
作者：
Sachs DH;Galli C
通讯作者：
Galli C
影响因子：
12.4
作者：
Mizuguchi, H;Xu, ZL;Hayakawa, T
通讯作者：
Hayakawa, T
影响因子：
2.9
作者：
Guan, Zheng-Bing;Dan, Wen-Bing;Zhang, Shuang-Quan
通讯作者：
Zhang, Shuang-Quan
DOI：
10.1186/1477-7827-1-82
发表时间：
2003-11-07
期刊：
Reproductive biology and endocrinology : RB&E
影响因子：
--
作者：
Lai, Liangxue;Prather, Randall S
通讯作者：
Prather, Randall S
影响因子：
4.5
作者：
de Felipe, P;Ryan, MD
通讯作者：
Ryan, MD