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The ability to induce the reprogramming of somatic mammalian cells to a pluripotent state by the forced expression of specific transcription factors has helped redefine the rules of cell fate and plasticity, as well as open possibilities for disease modeling, drug screening and regenerative medicine. Here, we hypothesized that the non-viral forced expression of the four originally discovered defined factors (OKSM) in adult mice could result in  in vivo  reprogramming of cells in the transfected tissue  in situ . We show that a single hydrodynamic tail-vein (HTV) injection of two plasmids encoding for  Oct3/4, Sox2, Klf4  and  c-Myc  respectively, are highly expressed in the liver tissue of Balb/C adult mice. Hallmark pluripotency markers were upregulated within 24–48 h after injection, followed by down-regulation of all major hepatocellular markers. Generation of transcriptionally reprogrammed cells  in vivo  was further confirmed by positive staining of liver tissue sections for all major pluripotency markers in Balb/C mice and the Nanog-GFP reporter transgenic strain (TNG-A) with concomitant upregulation of GFP expression  in situ . No signs of physiological or anatomical abnormalities or teratoma formation were observed in the liver examined up to 120 days. These findings indicate that virus-free expression of OKSM factors  in vivo  can transcriptionally reprogram cells  in situ  rapidly, efficiently and transiently, absent of host tissue damage or teratoma formation.

In Vivo Cell Reprogramming towards Pluripotency by Virus-Free Overexpression of Defined Factors