Gammaretroviral vector encoding a fluorescent marker to facilitate detection of reprogrammed human fibroblasts during iPSC generation

编码荧光标记的伽马逆转录病毒载体有助于在 iPSC 生成过程中检测重编程的人类成纤维细胞

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作者:Narasimhachar Srinivasakumar, Michail Zaboikin, Andrew M Tidball, Asad A Aboud, M Diana Neely, Kevin C Ess, Aaron B Bowman, Friedrich G Schuening

Abstract

Induced pluripotent stem cells (iPSCs) are becoming mainstream tools to study mechanisms of development and disease. They have a broad range of applications in understanding disease processes, in vitro testing of novel therapies, and potential utility in regenerative medicine. Although the techniques for generating iPSCs are becoming more straightforward, scientists can expend considerable resources and time to establish this technology. A major hurdle is the accurate determination of valid iPSC-like colonies that can be selected for further cloning and characterization. In this study, we describe the use of a gammaretroviral vector encoding a fluorescent marker, mRFP1, to not only monitor the efficiency of initial transduction but also to identify putative iPSC colonies through silencing of mRFP1 gene as a consequence of successful reprogramming.

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