Gene expression profile changes after short-activating RNA-mediated induction of endogenous pluripotency factors in human mesenchymal stem cells

Jon Voutila, Pål Sætrom, Paul Mintz, Guihua Sun, Jessica Alluin, John J. Rossi, Nagy A. Habib, Noriyuki Kasahara

Research output: Contribution to journalArticle

20 Scopus citations

Abstract

It is now recognized that small noncoding RNA sequences have the ability to mediate transcriptional activation of specific target genes in human cells. Using bioinformatics analysis and functional screening, we screened short-activating RNA (saRNA) oligonucleotides designed to target the promoter regions of the pluripotency reprogramming factors, Kruppel-like factor 4 (KLF4) and c-MYC. We identified KLF4 and c-MYC promoter-targeted saRNA sequences that consistently induced increases in their respective levels of nascent mRNA and protein expression in a time- and dose-dependent manner, as compared with scrambled sequence control oligonucleotides. The functional consequences of saRNA-induced activation of each targeted reprogramming factor were then characterized by comprehensively profiling changes in gene expression by microarray analysis, which revealed significant increases in mRNA levels of their respective downstream pathway genes. Notably, the microarray profile after saRNA-mediated induction of endogenous KLF4 and c-MYC showed similar gene expression patterns for stem cell- and cell cycle-related genes as compared with lentiviral vector-mediated overexpression of exogenous KLF4 and c-MYC transgenes, while divergent gene expression patterns common to viral vector-mediated transgene delivery were also noted. The use of promoter-targeted saRNAs for the activation of pluripotency reprogramming factors could have broad implications for stem cell research.

Original languageEnglish (US)
Article numbere35
Pages (from-to)e35
JournalMolecular Therapy - Nucleic Acids
Volume1
Issue number8
DOIs
StatePublished - 2012

Keywords

  • Gene expression profiling
  • Mesenchymal stem cells
  • Pluripotency genes
  • RNA activation
  • SaRNA

ASJC Scopus subject areas

  • Molecular Medicine
  • Drug Discovery

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