Reprogramming of mouse and human cells to pluripotency using mature microRNAs.

Induced pluripotent stem cells (iPSCs) can be generated from differentiated human and mouse somatic cells using transcription factors such as Oct4, Sox2, Klf4, and c-Myc. It is possible to augment the reprogramming process with chemical compounds, but issues related to low reprogramming efficiencies and, with a number of protocols, residual vector sequences, remain to be resolved. We show here that it is possible to reprogram mouse and human cells to pluripotency by direct transfection of mature double-stranded microRNAs (miRNAs). Our approaches use a combination of mir-200c plus mir-302 s and mir-369 s family miRNAs. Because this reprogramming method does not require vector-based gene transfer, it holds significant potential for biomedical research and regenerative medicine.

[1]  V. Ambros,et al.  The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14 , 1993, Cell.

[2]  T. Ichisaka,et al.  GENERATION OF GERMLINECOMPETENT INDUCED PLURIPOTENT STEM CELLS , 2007 .

[3]  Mudit Gupta,et al.  Highly efficient miRNA-mediated reprogramming of mouse and human somatic cells to pluripotency. , 2011, Cell stem cell.

[4]  Robert L. Judson,et al.  Embryonic stem cell–specific microRNAs promote induced pluripotency , 2009, Nature Biotechnology.

[5]  L. Lim,et al.  MicroRNA targeting specificity in mammals: determinants beyond seed pairing. , 2007, Molecular cell.

[6]  Wei Wang,et al.  piggyBac transposition reprograms fibroblasts to induced pluripotent stem cells , 2009, Nature.

[7]  E. Li,et al.  KDM1B is a histone H3K4 demethylase required to establish maternal genomic imprints , 2009, Nature.

[8]  S. Moon,et al.  Human embryonic stem cells express a unique set of microRNAs. , 2004, Developmental biology.

[9]  Alessandro Rosa,et al.  A regulatory circuitry comprised of miR‐302 and the transcription factors OCT4 and NR2F2 regulates human embryonic stem cell differentiation , 2011, The EMBO journal.

[10]  Robert Lanza,et al.  Generation of human induced pluripotent stem cells by direct delivery of reprogramming proteins. , 2009, Cell stem cell.

[11]  Donald C. Chang,et al.  Regulation of somatic cell reprogramming through inducible mir-302 expression , 2010, Nucleic acids research.

[12]  Wenjun Guo,et al.  Induction of pluripotent stem cells from primary human fibroblasts with only Oct4 and Sox2 , 2008, Nature Biotechnology.

[13]  Alexander Meissner,et al.  Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA. , 2010, Cell stem cell.

[14]  T. Ichisaka,et al.  Hypoxia enhances the generation of induced pluripotent stem cells. , 2009, Cell stem cell.

[15]  H. Redl,et al.  Vitamin C enhances the generation of mouse and human induced pluripotent stem cells. , 2010, Cell stem cell.

[16]  S. Kauppinen,et al.  Therapeutic Silencing of MicroRNA-122 in Primates with Chronic Hepatitis C Virus Infection , 2010, Science.

[17]  G. Goodall,et al.  The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1 , 2008, Nature Cell Biology.

[18]  G. Churchill,et al.  Characterization of human embryonic stem cell lines by the International Stem Cell Initiative , 2007, Nature Biotechnology.

[19]  Dong Wook Han,et al.  Generation of induced pluripotent stem cells using recombinant proteins. , 2009, Cell stem cell.

[20]  S. Yamanaka,et al.  Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors , 2006, Cell.

[21]  Shinya Yamanaka,et al.  Generation of Mouse Induced Pluripotent Stem Cells Without Viral Vectors , 2008, Science.

[22]  P. Sharp,et al.  Embryonic stem cell-specific MicroRNAs. , 2003, Developmental cell.

[23]  Wenjun Guo,et al.  Induction of pluripotent stem cells by defined factors is greatly improved by small-molecule compounds , 2008, Nature Biotechnology.

[24]  G. Ruvkun.  Molecular biology. Glimpses of a tiny RNA world. , 2001, Science.

[25]  Deepak M. Gupta,et al.  A nonviral minicircle vector for deriving human iPS cells , 2010, Nature Methods.

[26]  Shinsuke Yuasa,et al.  Generation of induced pluripotent stem cells from human terminally differentiated circulating T cells. , 2010, Cell stem cell.

[27]  T. Ichisaka,et al.  Generation of germline-competent induced pluripotent stem cells , 2007, Nature.

[28]  三吉 範克.  Defined factors induce reprogramming of gastrointestinal cancer cells , 2011 .

[29]  Megan F. Cole,et al.  Connecting microRNA Genes to the Core Transcriptional Regulatory Circuitry of Embryonic Stem Cells , 2008, Cell.

[30]  K. Woltjen,et al.  Virus free induction of pluripotency and subsequent excision of reprogramming factors , 2009, Nature.