Open AccessA Cassette System to Study Embryonic Stem Cell Differentiation by Inducible RNA Interference
Author(s) -
Wegmüller Daniel,
Raineri Ines,
Gross Brigitte,
Oakeley Edward J.,
Moroni Christoph
Publication year - 2007
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1634/stemcells.2006-0106
Subject(s) - biology , embryonic stem cell , embryoid body , microbiology and biotechnology , small hairpin rna , rna interference , induced pluripotent stem cell , cellular differentiation , stem cell , transcription factor , genetics , rna , gene
Although differentiation of pluripotent embryonic stem cells is restricted by a hierarchy of transcription factors, little is known about whether post‐transcriptional mechanisms similarly regulate early embryoid differentiation. We developed a system where small hairpin (sh)RNAs can be induced in embryonic stem (ES) cells from a defined locus following integration by Flp recombinase‐mediated DNA recombination. To verify the system, the key transcription factor Stat3, which maintains pluripotency, was downregulated by shRNA, and the expected morphological and biochemical markers of differentiation were observed. Induction of shRNA specific for the post‐transcriptional regulator Brf1 ( Zfp36L1 ) amplified the cardiac markers with strong stimulation of cardiomyocyte formation within embryoid bodies. These findings identify Brf1 as a novel potential regulator of cardiomyocyte formation and suggest that post‐transcriptional mechanisms are of importance to early development and, possibly, to regenerative medicine. The inducible RNA interference system presented here should also allow assignment of function for candidate genes with suspected roles in ES cell development. Disclosure of potential conflicts of interest is found at the end of this article.