State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, People's Republic of China; and.
Department of Human Genetics and.
Proc Natl Acad Sci U S A. 2014 Jul 22;111(29):10678-83. doi: 10.1073/pnas.1409933111. Epub 2014 Jul 7.
Induced pluripotent stem (iPS) cells hold great promise for regenerative medicine. To overcome potential problems associated with transgene insertions, efforts have been directed over the past several years to generate transgene-free iPS cells by using non-viral-vector approaches. To date, however, cells generated through such procedures have had problems producing reproductively competent animals, suggesting that their quality needed further improvement. Here we report the use of optimized assemblies of reprogramming factors and selection markers incorporated into single plasmids as nonintegrating episomes to generate germ-line-competent iPS cells. In particular, the pMaster12 episome can produce transgene-free iPS cells that, when grown in 2i medium, recapitulate good mouse ES cells, in terms of their competency for generating germ-line chimeras.
诱导多能干细胞（iPS 细胞）在再生医学方面具有巨大的应用潜力。为了克服与转基因插入相关的潜在问题，过去几年来，人们一直致力于通过非病毒载体方法来产生无转基因的 iPS 细胞。然而，迄今为止，通过这些方法产生的细胞在产生具有生殖能力的动物方面存在问题，这表明它们的质量需要进一步提高。在这里，我们报告了使用优化的重编程因子和选择标记物组装体，将其整合到单个质粒中作为非整合的附加体，以生成具有种系嵌合能力的 iPS 细胞。特别是，pMaster12 附加体可以产生无转基因的 iPS 细胞，当在 2i 培养基中生长时，这些细胞在产生种系嵌合体的能力方面与良好的小鼠 ES 细胞相当。
