|
|
|
|
|
| The Defined siRNAs Suppress Nanog and Sox2 Expressions in Mouse ES Cells |
| LEI Lei, DOU Lin , WANG Hua-yan |
| College of Veterinary Medicine/Shaanxi Center for Stem Cell Engineering and Technology, Northwest A&F University, |
|
|
|
摘要 Nanog, Oct4 and Sox2 are important transcription factors that are expressed in embryonic stem (ES) cells or embryoniccarcinoma (EC) cells, but in most cases they are absent in somatic cells. These factors play a key role to maintain embryonic stemcell self-renew and pluripotency. Down-regulation of Nanog and Sox2 gene expression can change multiple gene expressionpatterns and signal transduction pathways, and will initiate ES cell differentiation. This study was designed to select theefficient small interfering RNA (siRNA) fragments that inhibit Nanog and Sox2 gene expression in mouse J1 ES cells and P19 ECcells. Among synthesized siRNAs we screened out the siRNA N301 for Nanog and siRNA S720 for Sox2, which not only downregulatedof Nanog and Sox2 gene expression, but also interfered embryoid bodies formation. Our study provided the definedsiRNA fragments that could be used to investigate the epigenetic function of Nanog and Sox2 genes.
Abstract Nanog, Oct4 and Sox2 are important transcription factors that are expressed in embryonic stem (ES) cells or embryoniccarcinoma (EC) cells, but in most cases they are absent in somatic cells. These factors play a key role to maintain embryonic stemcell self-renew and pluripotency. Down-regulation of Nanog and Sox2 gene expression can change multiple gene expressionpatterns and signal transduction pathways, and will initiate ES cell differentiation. This study was designed to select theefficient small interfering RNA (siRNA) fragments that inhibit Nanog and Sox2 gene expression in mouse J1 ES cells and P19 ECcells. Among synthesized siRNAs we screened out the siRNA N301 for Nanog and siRNA S720 for Sox2, which not only downregulatedof Nanog and Sox2 gene expression, but also interfered embryoid bodies formation. Our study provided the definedsiRNA fragments that could be used to investigate the epigenetic function of Nanog and Sox2 genes.
|
|
Received: 19 August 2010
Accepted: 09 September 2011
|
| Fund: This work was supported by the National Natural ScienceFoundation of China (30871786) and the NationalBasic Research Program of China (973 Program, 2009CB941002). |
|
Corresponding Authors:
Correspondence WANG Hua-yan, Professor, Tel: +86-29-87080069, Fax: 86-29-87080068,E-mail: hhwang101@163.com
E-mail: hhwang101@163.com
|
| About author: LEI Lei, Ph D, Tel: +86-29-87080069, E-mail: friendly0722@126.com |
Cite this article:
LEI Lei, DOU Lin , WANG Hua-yan.
2011.
The Defined siRNAs Suppress Nanog and Sox2 Expressions in Mouse ES Cells. Journal of Integrative Agriculture, 10(9): 1475-1481.
|
| [1]Boyer L A, Lee T I, Cole M F, Johnstone S E, Levine S S, ZuckerJ P, Guenther M G, Kumar R M, Murray H L, Jenner R G,et al. 2005. Core transcriptional regulatory circuitry in humanembryonic stem cells. Cell, 122, 947-956.[2]Burdon T, Stracey C, Chambers I, Nichols J, Smith A. 1999.Suppression of SHP-2 and ERK signalling promotes selfrenewalof mouse embryonic stem cells. DevelopmentalBiology, 210, 30-43.[3]Chambers I, Colby D, Robertson M, Nichols J, Lee S, TweedieS, Smith A. 2003. Functional expression cloning of Nanog, apluripotency sustaining factor in embryonic stem cells. Cell,113, 643-655.[4]Chen S, Choo A B, Nai-Dy W, Heng-Phon T, Oh S K. 2007.Knockdown of Oct-4 or Sox-2 attenuates neurogenesis ofmouse embryonic stem cells. Stem Cell Developmental, 16,413-420.[5]Chew J L, Loh Y H, Zhang W, Chen X, Tam W L, Yeap L S, Li P,Ang Y S, Lim B, Robson P, et al. 2005, Reciprocaltranscriptional regulation of Pou5f1 and Sox2 via the Oct4/Sox2 complex in embryonic stem cells. Moecularl CellularBiology, 25, 6031-6046.[6]Dreesen O, Brivanlou A H. 2007. Signaling pathways in cancerand embryonic stem cells. Stem Cell Reviews, 3, 7-17.[7]Elbashir S M, Lendeckel W, Tuschl T. 2001. RNA interference ismediated by 21- and 22-nucleotide RNAs. Genes &Development, 15, 188-200.[8]Fire A, Xu S, Montgomery M K, Kostas S A, Driver S E, MelloC C. 1998. Potent and specific genetic interference by doublestrandedRNA in Caenorhabditis elegans. Nature, 391, 806-811.[9]Hannon G J. 2002. RNA interference. Nature, 418, 244-251.[10]Haruhiko S, Mikiko S. 2009. On the road to reading the RNAinterferencecode. Nature, 457, 396-404.[11]Hohenstein K A, Pyle A D, Chern J Y, Lock L F, Donovan P J.2008. Nucleofection mediates high-efficiency stable geneknockdown and transgene expression in human embryonicstem cells. Stem Cells, 26, 1436-1443.[12]Hyslop L, Stojkovic M, Armstrong L, Walter T, Stojkovic P,Przyborski S, Herbert M, Murdoch A, Strachan T, Lako M.2005. Downregulation of NANOG induces differentiationof human embryonic stem cells to extraembryonic lineages.Stem Cells, 23, 1035-1043.[13]Jackson A L, Linsley P S. 2004. Noise amidst the silence: offtargeteffects of siRNAs? Trends in Genetics, 20, 521-524.[14]Masui S, Nakatake Y, Toyooka Y, Shimosato D, Yagi R, TakahashiK, Okochi H, Okuda A, Matoba R, Sharov A A, et al. 2007.Pluripotency governed by Sox2 via regulation of Oct3/4expression in mouse embryonic stem cells. Nature CellBiology, 9, 625-635.[15]Mitsui K, Tokuzawa Y, Itoh H, Segawa K, Murakami M,Takahashi K, Maruyama M, Maeda M, Yamanaka S. 2003.The homeoprotein Nanog is required for maintenance ofpluripotency in mouse epiblast and ES cells. Cell, 113, 631-642.[16]Niwa H. 2001. Molecular mechanism to maintain stem cellrenewal of ES cells. Cell Structure and Function, 26, 137-148.[17]Niwa H, Burdon T, Chambers I, Smith A. 1998. Self-renewal ofpluripotent embryonic stem cells is mediated via activationof STAT3. Genes and Development, 12, 2048-2060.[18]Paddison P J, Caudy A A, Bernstein E, Hannon G J, Conklin DS. 2002. Short hairpin RNAs (shRNAs) induce sequencespecificsilencing in mammalian cells. Genes and Development,16, 948-958.[19]Pan G, Li J, Zhou Y, Zheng H, Pei D. 2006. A negative feedbackloop of transcription factors that controls stem cellpluripotency and self-renewal. FASEB Journal, 20, 1730-1732.[20]Pei D. 2009. Regulaiton of pluripotency and reprogramming bytranscription factors. The Journal of Biological Chemistry,284, 3365-3369.[21]Sato N, Meijer L, Skaltsounis L, Greengard P, Brivanlou A H.2004. Maintenance of pluripotency in human and mouseembryonic stem cells through activation of Wnt signaling bya pharmacological GSK-3-specific inhibitor. Nature Medicine,10, 55-63.[22]Smith A G. 2001. Embryo-derived stem cells: of mice and men.Annual Review of Cell and Developmental Biology, 17, 435-462.[23]Sumi T, Fujimoto Y, Nakatsuji N, Suemori H. 2004. STAT3 isdispensable for maintenance of self-renewal in nonhumanprimate embryonic stem cells. Stem Cells, 22, 861-872.[24]Takahashi K, Yamanaka S. 2006. Induction of pluripotent stemcells from mouse embryonice and adult fibroblast culturesby defined factors. Cell, 126, 663-676.[25]Yuko F, Stuart H, Orkin. 2009. Molecular regulation of the stateof embryonic stem cell. Stem Cell and Cancer, 33-60. |
| No Suggested Reading articles found! |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
