An Embryonic Stem Cell-Based System for Rapid Analysis of Transcriptional Enhancers
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CitationTsanov, Kaloyan M., Yuichi Nishi, Kevin A. Peterson, Jing Liu, Manfred Baetscher, and Andrew P. McMahon. Forthcoming. An embryonic stem cell-based system for rapid analysis of transcriptional enhancers. Genesis.
AbstractWith the growing use of genome-wide screens for cis-regulatory elements, there is a pressing need for platforms that enable fast and cost-effective experimental validation of identified hits in relevant developmental and tissue contexts. Here, we describe a murine embryonic stem cell (ESC)-based system that facilitates rapid analysis of putative transcriptional enhancers. Candidate enhancers are targeted with high efficiency to a defined genomic locus via recombinase-mediated cassette exchange. Targeted ESCs are subsequently differentiated in vitro into desired cell types, where enhancer activity is monitored by reporter gene expression. As a proof of principle, we analyzed a previously characterized, Sonic hedgehog (Shh)-dependent, V3 interneuron progenitor (pV3)-specific enhancer for the Nkx2.2 gene, and observed highly specific enhancer activity. Given the broad potential of ESCs to generate a spectrum of cell types, this system can serve as an effective platform for the characterization of gene regulatory networks controlling cell fate specification and cell function.
Citable link to this pagehttp://nrs.harvard.edu/urn-3:HUL.InstRepos:8715742
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