A medium‐scale assay for enhancer validation in amniotes. Issue 10 (12th August 2015)

• Record Type: Journal Article
• Authors: Chen, Jingchen; Streit, Andrea
• Is Part Of: Developmental dynamics. Volume 244:Issue 10(2015:Oct.)
• Page Start: 1291
• Page End: 1299
• Publication Date: 2015-08-12
• Subjects: Morphogenesis -- Periodicals; Anatomy -- Periodicals; Anatomie -- Périodiques; Biologie du développement -- Périodiques; 571.833
• DOI: 10.1002/dvdy.24306
• Languages: English
• ISSNs: 1058-8388

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <sec id="dvdy24306-sec-0001" sec-type="section"> <title>Background</title> <p>Enhancers are key elements to control gene expression in time and space and thus orchestrate gene function during development, homeostasis, and disease. Whole genome approaches and bioinformatic predictions have generated a tremendous pool of potential enhancers, however their spatiotemporal activity often remains to be validated in vivo. Despite recent progress in developing high throughput strategies for enhancer evaluation, these remain mainly restricted to invertebrates and in vitro cell culture.</p> </sec> <sec id="dvdy24306-sec-0002" sec-type="section"> <title>Results</title> <p>Here we design a medium‐scale method to validate potential enhancers in an amniote embryo, the chick. Using a unique barcode for different reporter vectors allows us to detect the activity of nine separate enhancers in a single embryo by one‐step RT‐PCR. The assay is sufficiently sensitive to expand its capacity further by generating additional barcoded vectors.</p> </sec> <sec id="dvdy24306-sec-0003" sec-type="section"> <title>Conclusions</title> <p>As a rapid, sensitive, and cost‐effective way to assess enhancer activity in an amniote vertebrate, this method provides a major advance and a useful alternative to the generation of transgenic animals. <italic>Developmental Dynamics 244:1291–1299, 2015</italic>. © 2015 The Authors. Developmental Dynamics published by Wiley Periodicals, Inc. on behalf of American Association of Anatomists</p> </sec> </abstract>