Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
권호 표지
DOI QR코드

DOI QR Code

Identification of Candidate Porcine miRNA-302/367 Cluster and Its Function in Somatic Cell Reprogramming

  • Son, Dong-Chan (Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute of Agriculture Life Sciences, Seoul National University) ;
  • Hwang, Jae Yeon (Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute of Agriculture Life Sciences, Seoul National University) ;
  • Lee, Chang-Kyu (Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute of Agriculture Life Sciences, Seoul National University)
  • Received : 2014.06.16
  • Accepted : 2014.06.23
  • Published : 2014.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

MicroRNAs (miRNAs) are approximately 22 nucleotides of small noncoding RNAs that control gene expression at the posttranscriptional level through translational inhibition and destabilization of their target mRNAs. The miRNAs are phylogenetically conserved and have been shown to be instrumental in a wide variety of key biological processes including cell cycle regulation, apoptosis, metabolism, imprinting, and differentiation. Recently, a paper has shown that expression of the miRNA-302/367 cluster expressed abundantly in mouse and human embryonic stem cells (ESCs) can directly reprogram mouse and human somatic cells to induced pluripotent stem cells (iPSCs) efficiently in the absence of any of the four factors, Oct4, Sox2, c-Myc, and Klf4. To apply this efficient method to porcine, we analyzed porcine genomic sequence containing predicted porcine miRNA-302/367 cluster through ENSEMBL database, generated a non-replicative episomal vector system including miRNA-302/367 cluster originated from porcine embryonic fibroblasts (PEF), and tried to make porcine iPSCs by transfection of the miRNA-302/367 cluster. Colonies expressing EGFP and forming compact shape were found, but they were not established as iPSC lines. Our data in this study show that pig miRNA-302/367 cluster could not satisfy requirement of PEF reprogramming conditions for pluripotency. To make pig iPSC lines by miRNA, further studies on the role of miRNAs in pluripotency and new trials of transfection with conventional reprogramming factors are needed.

Keywords

References

  1. Ambros V (2004): The functions of animal micro-RNAs. Nature 431:350-355. https://doi.org/10.1038/nature02871
  2. Anokye-Danso F, Trivedi CM, Juhr D, Gupta M, Cui Z, Tian Y, Zhang Y, Yang W, Gruber PJ, Epstein JA, Morrisey EE (2011): Highly efficient mi-RNA-mediated reprogramming of mouse and human somatic cells to pluripotency. Cell Stem Cell 8: 376-388. https://doi.org/10.1016/j.stem.2011.03.001
  3. Barroso-del Jesus A, Lucena-Aguilar G, Menendez P (2000): The miR-302-367 cluster as a potential stemness regulator in ESCs. Cell Cycle 8:394-398.
  4. Bartel DP (2004): MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116:281-297. https://doi.org/10.1016/S0092-8674(04)00045-5
  5. Hannon GJ (2002): RNA interference. Nature 418: 244-251. https://doi.org/10.1038/418244a
  6. Hotta A, Ellis J (2008): Retroviral vector silencing during iPS cell induction: an epigenetic beacon that signals distinct pluripotent states. J Cell Biochem 105:940-948. https://doi.org/10.1002/jcb.21912
  7. Hussein SM, Batada NN, Vuoristo S, Ching RW, Autio R, Narva E, Ng S, Sourour M, Hamalainen R, Olsson C, Lundin K, Mikkola M, Trokovic R, Peitz M, Brustle O, Bazett-Jones DP, Alitalo K, Lahesmaa R, Nagy A, Otonkoski T (2011): Copy number variation and selection during reprogramming to pluripotency. Nature 471:58-62. https://doi.org/10.1038/nature09871
  8. Lakshmipathy U, Love B, Goff LA, Jörnsten R, Graichen R, Hart RP, Chesnut JD (2007): MicroRNA expression pattern of undifferentiated and differentiated human embryonic stem cells. Stem Cells Dev 16:1003-1016. https://doi.org/10.1089/scd.2007.0026
  9. Laurent LC, Ulitsky I, Slavin I, Tran H, Schork A, Morey R, Lynch C, Harness JV, Lee S, Barrero MJ, Ku S, Martynova M, Semechkin R, Galat V, Gottesfeld J, Izpisua Belmonte JC, Murry C, Keirstead HS, Park HS, Schmidt U, Laslett AL, Muller FJ, Nievergelt CM, Shamir R, Loring JF (2011): Dynamic changes in the copy number of pluripotency and cell proliferation genes in human ESCs and iPSCs during reprogramming and time in culture. Cell Stem Cell 8:106-118. https://doi.org/10.1016/j.stem.2010.12.003
  10. Lister R, Pelizzola M, Kida YS, Hawkins RD, Nery JR, Hon G, Antosiewicz-Bourget J, O'Malley R, Castanon R, Klugman S, Downes M, Yu R, Stewart R, Ren B, Thomson JA, Evans RM, Ecker JR (2011): Hotspots of aberrant epigenomic reprogramming in human induced pluripotent stem cells. Nature 471: 68-73. https://doi.org/10.1038/nature09798
  11. Marson A, Levine SS, Cole MF, Frampton GM, Brambrink T, Johnstone S, Guenther MG, Johnston WK, Wernig M, Newman J, Calabrese JM, Dennis LM, Volkert TL, Gupta S, Love J, Hannett N, Sharp PA, Bartel DP, Jaenisch R, Young RA (2008): Connecting microRNA genes to the core transcriptional regulatory circuitry of embryonic stem cells. Cell 134:521-533. https://doi.org/10.1016/j.cell.2008.07.020
  12. Mayshar Y, Ben-David U, Lavon N, Biancotti JC, Yakir B, Clark AT, Plath K, Lowry WE, Benvenisty N (2010): Identification and classification of chromosomal aberrations in human Induced pluripotent stem cells. Cell Stem Cell 7:521-531. https://doi.org/10.1016/j.stem.2010.07.017
  13. Miyoshi N, Ishii H, Nagano H, Haraguchi N, Dewi DL, Kano Y, Nishikawa S, Tanemura M, Mimori K, Tanaka F, Saito T, Nishimura J, Takemasa I, Mizushima T, Ikeda M, Yamamoto H, Sekimoto M, Doki Y, Mori M (2011): Reprogramming of mouse and human cells to pluripotency using mature microRNAs. Cell Stem Cell 8:633-638. https://doi.org/10.1016/j.stem.2011.05.001
  14. Morin RD, O'Connor MD, Griffith M, Kuchenbauer F, Delaney A, Prabhu AL, Zhao Y, McDonald H, Zeng T, Hirst M, Eaves CJ, Marra MA (2008): Application of massively parallel sequencing to micro- RNA profiling and discovery in human embryonic stem cells. Genome Res 18:610-621. https://doi.org/10.1101/gr.7179508
  15. Pera MF (2011): Stem cells: The dark side of induced pluripotency. Nature 471:46-47. https://doi.org/10.1038/471046a
  16. Suh MR, Lee Y, Kim JY, Kim SK, Moon SH, Lee JY, Cha KY, Chung HM, Yoon HS, Moon SY, Kim VN, Kim KS (2004): Human embryonic stem cells express a unique set of microRNAs. Dev Biol 270: 488-498. https://doi.org/10.1016/j.ydbio.2004.02.019
  17. Takahashi K, Yamanaka S (2006): Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126: 663-676. https://doi.org/10.1016/j.cell.2006.07.024
  18. Warren L, Manos PD, Ahfeldt T, Loh YH, Li H, Lau F, Ebina W, Mandal PK, Smith ZD, Meissner A, Daley GQ, Brack AS, Collins JJ, Cowan C, Schlaeger TM, Rossi DJ (2010): Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified m- RNA. Cell Stem Cell 7:618-630. https://doi.org/10.1016/j.stem.2010.08.012
  19. Wong DJ, Segal E, Chang HY (2008): Stemness, cancer and cancer stem cells. Cell Cycle 7:3622-3624 https://doi.org/10.4161/cc.7.23.7104
  20. Yu J, Hu K, Smuga-Otto K, Tian S, Stewart R, Slukvin II, Thomson JA (2009): Human induced pluripotent stem cells free of vector and transgene sequences. Science 324:797-801. https://doi.org/10.1126/science.1172482