Identification of Differentially Expressed Genes in the Dicer 1 Knock-down Mouse Embryos using Microarray

  • Lee, Jae-Dal (Department of Pat Care, Hyejeon College) ;
  • Cui, Xiang-Shun (Department of Animal Sciences, Chungbuk National University)
  • Published : 2008.12.31

Abstract

Silencing of Dicer1 by siRNA did not inhibit development up to the blastocyst stage, but decreased expression of selected transcription factors, including Oct-4, Sox2 and Nanog, suggesting that Dicer1 gene expression is associated with differentiation processes at the blastocyst stage (Cui et al., 2007). In order to get insights into genes which may be linked with microRNA system, we compared gene expression profiles in Gapdh and Dicer1 siRNA-microinjected blastocysts using the Applied Biosystem microarray technology. Our data showed that 397 and 737 out of 16354 genes were up- and down-regulated, respectively, following siRNA microinjection (p<0.05), including 24 up- and 28 down-regulated transcription factors. Identification of genes that are preferentially expressed at particular Dicer1 knock down embryos provides insights into the complex gene regulatory networks that drive differentiation processes in embryos at blastocyst stage.

References

  1. Ambros V, Bartel B, Bartel DP, Burge CB, Carrington JC, Chen X, Dreyfuss G, Eddy SR, Griffinths-Jones S, Marshall M (2003): A uniform system for microRNA annotation. RNA 9:277-279 https://doi.org/10.1261/rna.2183803
  2. Benjamini Y, Hochberg Y (1995): Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc Ser B: Methodological 57:289-300
  3. Bernstein E, Caudy AA, Hammond SM, Hannon GJ (2001): Role for a bidentate ribonuclease in the initiation step of RNA interference. Nature 409:363-366 https://doi.org/10.1038/35053110
  4. Cui XS, Shen XH, Kim NH (2007): Dicer1 expression in preimplantation mouse embryos: Involvement of Oct3/4 transcription at the blastocyst stage, Biochem Biophys Res Commun 352:231-236 https://doi.org/10.1016/j.bbrc.2006.11.009
  5. Hamatani T, Daikoku T, Wang H, Matsumoto H, Carter MG, Ko MS, Dey SK (2004): Global gene expression analysis identifies molecular pathways distinguishing blastocyst dormancy and activation. Proc Natl Acad Sci USA 101:10326-10331 https://doi.org/10.1073/pnas.0402597101
  6. Kanellopoulou C, Muljo SA, Kung AL, Ganesan S, Drapkin R, Jenuwein T, Livingston DM, Rajewsky K (2005): Dicer-deficient mouse embryonic stem cells are defective in differentiation and centromeric silencing. Genes Dev 19:489-501 https://doi.org/10.1101/gad.1248505
  7. Livak KJ, Schmittgen TD (2001): Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)). Method 25: 402-408 https://doi.org/10.1006/meth.2001.1262
  8. SAS User's Guide: Statistics, Version 5 (1985): Cary, NC, SAS
  9. Steel RGD, Torrie JH (1980): Principles and Procedures of Statistics, McGraw Hill Book Co., New York, 1980
  10. Tanaka TS, Ko MSH (2004): A global view of gene expression in the preimplantation mouse embryo: Morula versus blastocyst. Obstetrics & Gynecology and Reproductive Biology 115S:S85-S91
  11. Wang QT, Piotrowska K, Ciemerych MA, Milenkovic L, Scott MP, Davis RW, Zernicka-Goetz M (2004): A genome-wide study of gene activity reveals developmental signaling pathways in the preimplantation mouse embryo. Dev Cell 6:133-144 https://doi.org/10.1016/S1534-5807(03)00404-0
  12. Whitworth KM, Agca C, Kim JG, Patel RV, Springer GK, Bivens NJ, Forrester LJ, Mathialagan N, Green JA, Prather RS (2005): Transcriptional profiling of pig embryogenesis by using a 15-K member unigene set specific for pig reproductive tissues and embryos. Biol Reprod 72:1437-1451 https://doi.org/10.1095/biolreprod.104.037952
  13. Zeng F, Baldwin DA, Schultz RM (2004): Transcript profiling during preimplantation mouse development. Dev Biol 272:483-496 https://doi.org/10.1016/j.ydbio.2004.05.018