Brief Report: Impaired Cell Reprogramming in Nonhomologous End Joining Deficient Cells

摘要: Although there is an increasing interest in defining the role of DNA damage response mechanisms cell reprogramming, relevance proteins participating nonhomologous end joining (NHEJ), a major mechanism double-strand breaks repair, this process remains to be investigated. Herein, we present data related reprogramming primary mouse embryonic fibroblasts (MEF) from severe combined immunodeficient (Scid) mice defective DNA-PKcs, key protein for NHEJ. Reduced numbers induced pluripotent stem (iPSC) colonies were generated Scid cells using lentiviral vectors (LV), being efficiency fourfold sevenfold lower than that observed wt cells. Moreover, these iPSC-like clones prematurely lost or differentiated spontaneously. While mutation neither reduce proliferation rate nor transduction efficacy transduced with LV, both expression SA-β-Gal and P16/INK4a senescence markers highly increased versus MEFs during process, accounting reduced MEFs. The use improved Sleeping Beauty transposon/transposase systems allowed us, however, isolate DNA-PKcs-deficient iPSCs which preserved their parental genotype hypersensitivity ionizing radiation. This new disease-specific iPSC model would useful understand physiological consequences DNA-PKcs development help improve current gene therapy strategies disease. STEM Cells 2013;31:1726–1730