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Homologous recombination DNA repair genes play a critical role in reprogramming to a pluripotent state.

Cell reports (2013-03-13)
Federico González, Daniela Georgieva, Fabio Vanoli, Zhong-Dong Shi, Matthias Stadtfeld, Thomas Ludwig, Maria Jasin, Danwei Huangfu
ABSTRACT

Induced pluripotent stem cells (iPSCs) hold great promise for personalized regenerative medicine. However, recent studies show that iPSC lines carry genetic abnormalities, suggesting that reprogramming may be mutagenic. Here, we show that the ectopic expression of reprogramming factors increases the level of phosphorylated histone H2AX, one of the earliest cellular responses to DNA double-strand breaks (DSBs). Additional mechanistic studies uncover a direct role of the homologous recombination (HR) pathway, a pathway essential for error-free repair of DNA DSBs, in reprogramming. This role is independent of the use of integrative or nonintegrative methods in introducing reprogramming factors, despite the latter being considered a safer approach that circumvents genetic modifications. Finally, deletion of the tumor suppressor p53 rescues the reprogramming phenotype in HR-deficient cells primarily through the restoration of reprogramming-dependent defects in cell proliferation and apoptosis. These mechanistic insights have important implications for the design of safer approaches to creating iPSCs.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
JumpStart Taq DNA Polymerase, with MgCl2
Sigma-Aldrich
Anti-phospho-Histone H2A.X (Ser139) Antibody, clone JBW301, clone JBW301, Upstate®, from mouse