An Embryonic Stem Cell-Based System for Rapid Analysis of Transcriptional Enhancers

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An Embryonic Stem Cell-Based System for Rapid Analysis of Transcriptional Enhancers

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Title: An Embryonic Stem Cell-Based System for Rapid Analysis of Transcriptional Enhancers
Author: Tsanov, Kaloyan M; Nishi, Yuichi; Peterson, Kevin A; Liu, Jing; Baetscher, Manfred; McMahon, Andrew P.

Note: Order does not necessarily reflect citation order of authors.

Citation: Tsanov, 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.
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Abstract: 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.
Published Version: doi:10.1002/dvg.20820
Terms of Use: This article is made available under the terms and conditions applicable to Open Access Policy Articles, as set forth at http://nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#OAP
Citable link to this page: http://nrs.harvard.edu/urn-3:HUL.InstRepos:8715742

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  • FAS Scholarly Articles [7362]
    Peer reviewed scholarly articles from the Faculty of Arts and Sciences of Harvard University
 
 

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