Induction of NANOG expression by targeting promoter sequence with small activating RNA antagonizes retinoic acid-induced differentiation.

The Biochemical Journal
Xiaoling WangLong-Cheng Li

Abstract

RNAa (RNA activation) is a mechanism by which small dsRNA (double-stranded RNA), termed saRNA (small activating RNA), target promoter sequences to induce gene expression. This technique represents a novel approach to gene overexpression without the use of exogenous DNA. In the present study, we investigated whether RNAa can modulate expression of the development-related gene NANOG and manipulate cell fate. Using a lentivirus-based reporter system as a screening tool, we identified synthetic saRNAs that stimulate NANOG expression in human NCCIT embryonic carcinoma cells. Mismatch mutations to saRNA duplexes define sequence requirement for gene activation. Functional analysis of NANOG induction reveals saRNA treatment predictably modulates the expression of several known downstream target genes, including FOXH1 (forkhead box H1), REST (RE1-silencing transcription factor), OCT4 (octamer-binding protein 4) and REX1 (reduced expression protein 1). Treatment with RA (retinoic acid) triggers NCCIT cell differentiation, reducing NANOG and OCT4 expression and up-regulating several neural markers [i.e. ASCL1 (achaete-scute complex homologue 1), NEUROD1 (neuronal differentiation 1) and PAX6 (paired box 6)]. However, co-treatment with saRN...Continue Reading

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Citations

Oct 12, 2013·PloS One·Barbara HämmerleJaime Font de Mora
Nov 8, 2013·Functional & Integrative Genomics·Sadegh Azimzadeh JamalkandiAli Masoudi-Nejad
Apr 16, 2013·The International Journal of Biochemistry & Cell Biology·Kai YangLi-Ping Xie
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Feb 24, 2016·Cell Research·Victoria PortnoyLong-Cheng Li
Sep 10, 2013·Wiley Interdisciplinary Reviews. Developmental Biology·Mohammad Zeeshan OzairAli H Brivanlou
Jun 16, 2015·Frontiers in Nutrition·Rosanna H E Krakowsky, Trygve O Tollefsbol
Oct 24, 2013·Epigenetics : Official Journal of the DNA Methylation Society·Long-Cheng Li
Nov 5, 2013·Epigenetics : Official Journal of the DNA Methylation Society·Alan L Jiao, Frank J Slack
Sep 28, 2014·Cancer Letters·Lin ZhengHao Zhang
Sep 19, 2016·Journal of Experimental & Clinical Cancer Research : CR·Huan-Lei WuZhang-Qun Ye
Jul 13, 2017·Antioxidants & Redox Signaling·Nihay Laham-KaramSeppo Ylä-Herttuala
Aug 23, 2012·Nucleic Acid Therapeutics·Mika KosakaLong-Cheng Li
Sep 14, 2018·Environmental Science and Pollution Research International·Ravi Prakash YadavNachimuthu Senthil Kumar
Mar 20, 2016·Journal of Biomolecular Screening·Bin WangYunZhang Hu

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Methods Mentioned

BETA
transfection
FACS
flow cytometry
transfections

Software Mentioned

WinMDI
RNAa

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