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Screening for Epigenetic Target Genes that Enhance Reprogramming Using Lentiviral-Delivered shRNA

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Adipose-Derived Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 702))

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Abstract

Small molecules will need to be identified and/or developed that target protein classes limiting reprogramming efficiency. A specific class of proteins includes epigenetic regulators that silence, or minimize expression, of pluripotency genes in differentiated cells. To better understand the role of specific epigenetic modulators in reprogramming, we have used shRNA delivered by lentivirus to assess the significance of individual epi-proteins in reprogramming pluripotent gene expression.

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References

  1. Lowry, W.E., et al., Generation of human induced pluripotent stem cells from dermal fibroblasts. Proceedings of the National Academy of Sciences of the United States of America, 2008. 105: p. 2883–2888.

    Article  PubMed  Google Scholar 

  2. Maherali, N., et al., Directly reprogrammed fibroblasts show global epigenetic remodeling and widespread tissue contribution. Cell Stem Cell, 2007. 1(1): p. 55–70.

    Article  PubMed  CAS  Google Scholar 

  3. Meissner, A., M. Wernig, and R. Jaenisch, Direct reprogramming of genetically unmodified fibroblasts into pluripotent stem cells. Nature Biotechnology, 2007. 25(10): p. 1171–1181.

    Article  CAS  Google Scholar 

  4. Okita, K., T. Ichisaka, and S. Yamanaka, Generation of germline-competent induced pluripotent stem cells. Nature, 2007. 448(7151): p. 313–317.

    Article  PubMed  CAS  Google Scholar 

  5. Okita, K., M. Nakagawa, H. Hyenjong, T. Ichisaka, and S. Yamanaka, Generation of mouse induced pluripotent stem cells without viral vectors. Science, 2008. 322(5903): p. 949–953.

    Article  PubMed  CAS  Google Scholar 

  6. Park, I., P.H. Lerou, R. Zhao, H. Huo, and G.Q. Daley, Generation of human-induced pluripotent stem cells. Nature Protocols, 2008. 3(7): p. 1180–1186.

    Article  PubMed  CAS  Google Scholar 

  7. Park, I.H., N. Arora, H. Huo, N. Maherali, T. Ahfeldt, A. Shimamura, C. Cowan, M.W. Lensch, K. Hochedlinger, and G.Q. Daley, Disease-specific induced pluripotent stem cells. Cell, 2008. 134: p. 1–10.

    Article  CAS  Google Scholar 

  8. Takahashi, K. and S. Yamanaka, Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell, 2006. 126(4): p. 663–676.

    Article  PubMed  CAS  Google Scholar 

  9. Takahashi, K., et al., Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell, 2007. 131: p. 861–872.

    Article  PubMed  CAS  Google Scholar 

  10. Aoi, T., K. Yae, M. Nakagawa, T. Ichisaka, K. Okita, K. Takahashi, T. Chiba, and S. Yamanaka, Generation of pluripotent stem cells from adult mouse liver and stomach cells. Science, 2008. 321(5889): p. 1–4.

    Article  CAS  Google Scholar 

  11. Wernig, M., et al., In vitro reprogramming of fibroblasts into a pluripotent ES-cell-like state. Nature, 2007. 448(7151): p. 318–324.

    Article  PubMed  CAS  Google Scholar 

  12. Yu, J., M.A. Vodyanik, K.S. Otto, J.A. Bourget, J.L. Frane, S. Tian, J. Nie, G.A. Jonsdottir, V. Ruotti, R. Stewart, I.I. Slukvin, and J.A. Thomson, Induced pluripotent stem cell lines derived from human somatic cells. Science, 2007. 318(5858): p. 1917–1920.

    Article  PubMed  CAS  Google Scholar 

  13. Mikkelsen, T.S., J. Hanna, X. Zhang, M. Ku, M. Wernig, P. Schorderet, B.E. Bernstein, R. Jaenisch, E.S. Lander, and A. Meissner, Dissecting direct reprogramming through integrative genomic analysis. Nature, 2008. 454(7200): p. 49–55.

    Article  PubMed  CAS  Google Scholar 

  14. Huangfu, D., R. Maehr, W. Guo, A. Eijkelenboom, M. Snitow, A.E. Chen, and D.A. Melton, Induction of pluripotent stem cells by defined factors is greatly improved by small-molecule compounds. Nature Biotechnology, 2008. 26(7): p. 795–797.

    Article  PubMed  CAS  Google Scholar 

  15. Huangfu, D., K. Osafune, R. Maehr, W. Guo, A. Eijkelenboom, S. Chen, W. Muhlestein, and D.A. Melton, Induction of pluripotent stem cells from primary human fibroblasts with only Oct4 and Sox2. Nature Biotechnology, 2008. 26(11): p. 1269–1275.

    Article  PubMed  CAS  Google Scholar 

  16. Feng, B., J. Jiang, P. Kraus, J-H. Ng, J-C.D. Heng, Y. Chan, L-P. Yaw, W. Zhang, Y-H. Loh, J. Han, V.B. Vega, V.C. Rataboul, B. Lim, T. Lufkin, and H-H. Ng, Reprogramming of fibroblasts into induced pluripotent stem cells with orphan nuclear receptor Esrrb. Nature Cell Biology, 2009. 11(2): p. 197–203.

    Article  PubMed  CAS  Google Scholar 

  17. Lyssiotis, C.A., R.K. Foreman, J. Staerk, M. Garcia, D. Mathur, S. Markoulaki, L.L. Lairson, J. Hanna, B.D. Charette, L.C. Bouchez, M. Bollong, C. Kunick, C.Y. Cho, A. Brinker, P.G. Schultz, and R. Jaenisch, Reprogramming of murine fibroblasts to induced pluripotent stem cells with chemical complementation of Klf4. Proceedings of the National Academy of Sciences of the United States of America, 2009. 106(22): p. 8912–8917.

    Article  PubMed  Google Scholar 

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Author information

Authors and Affiliations

  1. Nuclear Reprogramming and Epigenetics Laboratory, NuPotential, Inc., Louisiana Emerging Technology Center, Baton Rouge, LA, USA

    Jong S. Rim, Karen Strickler, Christian Barnes, Lettie Harkins, Jaroslaw Staszkiewicz, Rachel Power & Kenneth J. Eilertsen

  2. Nuclear Reprogramming and Epigenetics Laboratory, Pennington Biomedical Research Center, Louisiana State University System, Baton Rouge, LA, USA

    Kenneth J. Eilertsen

Authors
  1. Jong S. Rim
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  2. Karen Strickler
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  3. Christian Barnes
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  4. Lettie Harkins
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  5. Jaroslaw Staszkiewicz
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  6. Rachel Power
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  7. Kenneth J. Eilertsen
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Corresponding author

Correspondence to Kenneth J. Eilertsen .

Editor information

Editors and Affiliations

  1. , Stem Cell Biology, Pennington Biomedical Research Center, Perkins Rd. 6400, Baton Rouge, 70808, Louisiana, USA

    Jeffrey M. Gimble

  2. School of Medicine, Department of Pharmacology, Tulane University, Tulane Ave. 1430, New Orleans, 70112-2699, Louisiana, USA

    Bruce A. Bunnell

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© 2011 Springer Science+Business Media, LLC

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Rim, J.S. et al. (2011). Screening for Epigenetic Target Genes that Enhance Reprogramming Using Lentiviral-Delivered shRNA. In: Gimble, J., Bunnell, B. (eds) Adipose-Derived Stem Cells. Methods in Molecular Biology, vol 702. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61737-960-4_22

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  • DOI: https://doi.org/10.1007/978-1-61737-960-4_22

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61737-959-8

  • Online ISBN: 978-1-61737-960-4

  • eBook Packages: Springer Protocols

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