Retroviral vector silencing during iPS cell induction: an epigenetic beacon that signals distinct pluripotent states

A Hotta, J Ellis - Journal of cellular biochemistry, 2008 - Wiley Online Library
Journal of cellular biochemistry, 2008Wiley Online Library
Retroviral vectors are transcriptionally silent in pluripotent stem cells. This feature has been
potently applied in studies that reprogram somatic cells into induced pluripotent stem (iPS)
cells. By delivering the four Yamanaka factors in retroviral vectors, high expression is
obtained in fibroblasts to induce the pluripotent state. Partial reprogramming generates
Class I iPS cells that express the viral transgenes and endogenous pluripotency genes. Full‐
reprogramming in Class II iPS cells silences the vectors as the endogenous genes maintain …
Abstract
Retroviral vectors are transcriptionally silent in pluripotent stem cells. This feature has been potently applied in studies that reprogram somatic cells into induced pluripotent stem (iPS) cells. By delivering the four Yamanaka factors in retroviral vectors, high expression is obtained in fibroblasts to induce the pluripotent state. Partial reprogramming generates Class I iPS cells that express the viral transgenes and endogenous pluripotency genes. Full‐reprogramming in Class II iPS cells silences the vectors as the endogenous genes maintain the pluripotent state. Thus, retroviral vector silencing serves as a beacon marking the fully reprogrammed pluripotent state. Here we review known silencer elements, and the histone modifying and DNA methylation pathways, that silence retroviral and lentiviral vectors in pluripotent stem cells. Both retroviral and lentiviral vectors are influenced by position effects and often exhibit variegated expression. The best vector designs facilitate full‐reprogramming and subsequent retroviral silencing, which is required for directed‐differentiation. Current retroviral reprogramming methods can be immediately applied to create patient‐specific iPS cell models of human disease, however, future clinical applications will require novel chemical or other reprogramming methods that reduce or eliminate the integrated vector copy number load. Nevertheless, retroviral vectors will continue to play an important role in genetically correcting patient iPS cell models. We anticipate that novel pluripotent‐specific reporter vectors will select for isolation of high quality human iPS cell lines, and select against undifferentiated pluripotent cells during regenerative medicine to prevent teratoma formation after transplantation. J. Cell. Biochem. 105: 940–948, 2008. © 2008 Wiley‐Liss, Inc.
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