Generation of an Enhancer-Trapping Vector for Insertional Mutagenesis in Zebrafish

Enhancer trapping (ET) is a powerful approach to establish tissue- or cell-specific reporters and identify expression patterns of uncharacterized genes. Although a number of enhancer-trapping vectors have been developed and a large library of fish lines with distinct tissue- or cell-specific express...

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Main Authors:	Liu, Chunyan, Song, Guili, Mao, Lin, Long, Yong, Li, Qing, Cui, Zongbin
Format:	Online
Language:	English
Published:	Public Library of Science 2015
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4593583/

id	pubmed-4593583
recordtype	oai_dc
spelling	pubmed-45935832015-10-14 Generation of an Enhancer-Trapping Vector for Insertional Mutagenesis in Zebrafish Liu, Chunyan Song, Guili Mao, Lin Long, Yong Li, Qing Cui, Zongbin Research Article Enhancer trapping (ET) is a powerful approach to establish tissue- or cell-specific reporters and identify expression patterns of uncharacterized genes. Although a number of enhancer-trapping vectors have been developed and a large library of fish lines with distinct tissue- or cell-specific expression of reporter genes have been generated, the specificity and efficiency of trapping vectors need to be improved because of the bias interaction of minimal promoters with genomic enhancers. Accordingly, we generated an enhancer-trapping vector pTME that contains a minimal mouse metallothionein gene (mMTI) promoter upstream of EGFP reporter. In the first round of screening, twelve zebrafish lines that carry a single copy of ET cassettes were characterized to have tissue- or cell-specific EGFP expression. One of the highly conserved noncoding elements near an insertion site of trapping cassettes was characterized as an enhancer that can specifically regulate the expression of EGFP in cells of the central nervous system. In addition, the pTME vector contains a mutation-cassette that is able to effectively block the transcription of an endogenous gene in an ET line with ubiquitous EGFP expression. Thus, the pTME vector can be used as an alternative tool for both enhancer trapping and mutagenesis across a target genome. Public Library of Science 2015-10-05 /pmc/articles/PMC4593583/ /pubmed/26436547 http://dx.doi.org/10.1371/journal.pone.0139612 Text en © 2015 Liu et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Liu, Chunyan Song, Guili Mao, Lin Long, Yong Li, Qing Cui, Zongbin
spellingShingle	Liu, Chunyan Song, Guili Mao, Lin Long, Yong Li, Qing Cui, Zongbin Generation of an Enhancer-Trapping Vector for Insertional Mutagenesis in Zebrafish
author_facet	Liu, Chunyan Song, Guili Mao, Lin Long, Yong Li, Qing Cui, Zongbin
author_sort	Liu, Chunyan
title	Generation of an Enhancer-Trapping Vector for Insertional Mutagenesis in Zebrafish
title_short	Generation of an Enhancer-Trapping Vector for Insertional Mutagenesis in Zebrafish
title_full	Generation of an Enhancer-Trapping Vector for Insertional Mutagenesis in Zebrafish
title_fullStr	Generation of an Enhancer-Trapping Vector for Insertional Mutagenesis in Zebrafish
title_full_unstemmed	Generation of an Enhancer-Trapping Vector for Insertional Mutagenesis in Zebrafish
title_sort	generation of an enhancer-trapping vector for insertional mutagenesis in zebrafish
description	Enhancer trapping (ET) is a powerful approach to establish tissue- or cell-specific reporters and identify expression patterns of uncharacterized genes. Although a number of enhancer-trapping vectors have been developed and a large library of fish lines with distinct tissue- or cell-specific expression of reporter genes have been generated, the specificity and efficiency of trapping vectors need to be improved because of the bias interaction of minimal promoters with genomic enhancers. Accordingly, we generated an enhancer-trapping vector pTME that contains a minimal mouse metallothionein gene (mMTI) promoter upstream of EGFP reporter. In the first round of screening, twelve zebrafish lines that carry a single copy of ET cassettes were characterized to have tissue- or cell-specific EGFP expression. One of the highly conserved noncoding elements near an insertion site of trapping cassettes was characterized as an enhancer that can specifically regulate the expression of EGFP in cells of the central nervous system. In addition, the pTME vector contains a mutation-cassette that is able to effectively block the transcription of an endogenous gene in an ET line with ubiquitous EGFP expression. Thus, the pTME vector can be used as an alternative tool for both enhancer trapping and mutagenesis across a target genome.
publisher	Public Library of Science
publishDate	2015
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4593583/
_version_	1613483585963032576

Generation of an Enhancer-Trapping Vector for Insertional Mutagenesis in Zebrafish

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