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An embryonic stem cell‐based system for rapid analysis of transcriptional enhancers
Author(s) -
Tsanov Kaloyan M.,
Nishi Yuichi,
Peterson Kevin A.,
Liu Jing,
Baetscher Manfred,
McMahon Andrew P.
Publication year - 2012
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/dvg.20820
Subject(s) - enhancer , biology , embryonic stem cell , enhancer trap , reporter gene , sonic hedgehog , enhancer rnas , progenitor cell , cell fate determination , microbiology and biotechnology , genetics , computational biology , gene , stem cell , gene expression , transcription factor
With 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. genesis 50: 443–450, 2012. © 2011 Wiley Periodicals, Inc.