title: Gammaretroviral vector encoding a fluorescent marker to facilitate detection of reprogrammed human fibroblasts during iPSC generation.
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.
authors.affiliation: Division of Hematology/Oncology, Department of Internal Medicine, Saint Louis University , Saint Louis, MO , USA.
authors.forename: Narasimhachar
authors.initials: N
authors.lastname: Srinivasakumar
authors.affiliation: Division of Hematology/Oncology, Department of Internal Medicine, Saint Louis University , Saint Louis, MO , USA.
authors.forename: Michail
authors.initials: M
authors.lastname: Zaboikin
authors.affiliation: Vanderbilt University Medical Center, Department of Neurology, Vanderbilt Kennedy Center for Research on Human Development , Nashville, TN , USA.
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Tidball
authors.affiliation: Vanderbilt University Medical Center, Department of Neurology, Vanderbilt Kennedy Center for Research on Human Development , Nashville, TN , USA.
authors.forename: Asad A
authors.initials: AA
authors.lastname: Aboud
authors.affiliation: Vanderbilt University Medical Center, Department of Neurology, Vanderbilt Kennedy Center for Research on Human Development , Nashville, TN , USA.
authors.forename: M Diana
authors.initials: MD
authors.lastname: Neely
authors.affiliation: Vanderbilt University Medical Center, Department of Neurology, Vanderbilt Kennedy Center for Research on Human Development , Nashville, TN , USA.
authors.forename: Kevin C
authors.initials: KC
authors.lastname: Ess
authors.affiliation: Vanderbilt University Medical Center, Department of Neurology, Vanderbilt Kennedy Center for Research on Human Development , Nashville, TN , USA.
authors.forename: Aaron B
authors.initials: AB
authors.lastname: Bowman
authors.affiliation: Division of Hematology/Oncology, Department of Internal Medicine, Saint Louis University , Saint Louis, MO , USA.
authors.forename: Friedrich G
authors.initials: FG
authors.lastname: Schuening
keywords: Detection; Gammaretrovirus; IPSC; Induced pluripotent stem cells; Reprogramming; Silencing