Quantitative spatial and temporal assessment of regulatory element activity in zebrafish

Mutations or genetic variation in noncoding regions of the genome harbouring cis-regulatory elements (CREs), or enhancers, have been widely implicated in human disease and disease risk. However, our ability to assay the impact of these DNA sequence changes on enhancer activity is currently very limi...

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Autores principales: Shipra Bhatia, Dirk Jan Kleinjan, Kirsty Uttley, Anita Mann, Nefeli Dellepiane, Wendy A Bickmore
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Lenguaje:EN
Publicado: eLife Sciences Publications Ltd 2021
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Acceso en línea:https://doaj.org/article/cabea42dcfad455facb02d1a512d8384
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spelling oai:doaj.org-article:cabea42dcfad455facb02d1a512d83842021-11-19T11:55:32ZQuantitative spatial and temporal assessment of regulatory element activity in zebrafish10.7554/eLife.656012050-084Xe65601https://doaj.org/article/cabea42dcfad455facb02d1a512d83842021-11-01T00:00:00Zhttps://elifesciences.org/articles/65601https://doaj.org/toc/2050-084XMutations or genetic variation in noncoding regions of the genome harbouring cis-regulatory elements (CREs), or enhancers, have been widely implicated in human disease and disease risk. However, our ability to assay the impact of these DNA sequence changes on enhancer activity is currently very limited because of the need to assay these elements in an appropriate biological context. Here, we describe a method for simultaneous quantitative assessment of the spatial and temporal activity of wild-type and disease-associated mutant human CRE alleles using live imaging in zebrafish embryonic development. We generated transgenic lines harbouring a dual-CRE dual-reporter cassette in a pre-defined neutral docking site in the zebrafish genome. The activity of each CRE allele is reported via expression of a specific fluorescent reporter, allowing simultaneous visualisation of where and when in development the wild-type allele is active and how this activity is altered by mutation.Shipra BhatiaDirk Jan KleinjanKirsty UttleyAnita MannNefeli DellepianeWendy A BickmoreeLife Sciences Publications Ltdarticleenhancergene regulationlive imagingnoncoding genometransgenic assayhuman diseaseMedicineRScienceQBiology (General)QH301-705.5ENeLife, Vol 10 (2021)
institution DOAJ
collection DOAJ
language EN
topic enhancer
gene regulation
live imaging
noncoding genome
transgenic assay
human disease
Medicine
R
Science
Q
Biology (General)
QH301-705.5
spellingShingle enhancer
gene regulation
live imaging
noncoding genome
transgenic assay
human disease
Medicine
R
Science
Q
Biology (General)
QH301-705.5
Shipra Bhatia
Dirk Jan Kleinjan
Kirsty Uttley
Anita Mann
Nefeli Dellepiane
Wendy A Bickmore
Quantitative spatial and temporal assessment of regulatory element activity in zebrafish
description Mutations or genetic variation in noncoding regions of the genome harbouring cis-regulatory elements (CREs), or enhancers, have been widely implicated in human disease and disease risk. However, our ability to assay the impact of these DNA sequence changes on enhancer activity is currently very limited because of the need to assay these elements in an appropriate biological context. Here, we describe a method for simultaneous quantitative assessment of the spatial and temporal activity of wild-type and disease-associated mutant human CRE alleles using live imaging in zebrafish embryonic development. We generated transgenic lines harbouring a dual-CRE dual-reporter cassette in a pre-defined neutral docking site in the zebrafish genome. The activity of each CRE allele is reported via expression of a specific fluorescent reporter, allowing simultaneous visualisation of where and when in development the wild-type allele is active and how this activity is altered by mutation.
format article
author Shipra Bhatia
Dirk Jan Kleinjan
Kirsty Uttley
Anita Mann
Nefeli Dellepiane
Wendy A Bickmore
author_facet Shipra Bhatia
Dirk Jan Kleinjan
Kirsty Uttley
Anita Mann
Nefeli Dellepiane
Wendy A Bickmore
author_sort Shipra Bhatia
title Quantitative spatial and temporal assessment of regulatory element activity in zebrafish
title_short Quantitative spatial and temporal assessment of regulatory element activity in zebrafish
title_full Quantitative spatial and temporal assessment of regulatory element activity in zebrafish
title_fullStr Quantitative spatial and temporal assessment of regulatory element activity in zebrafish
title_full_unstemmed Quantitative spatial and temporal assessment of regulatory element activity in zebrafish
title_sort quantitative spatial and temporal assessment of regulatory element activity in zebrafish
publisher eLife Sciences Publications Ltd
publishDate 2021
url https://doaj.org/article/cabea42dcfad455facb02d1a512d8384
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AT kirstyuttley quantitativespatialandtemporalassessmentofregulatoryelementactivityinzebrafish
AT anitamann quantitativespatialandtemporalassessmentofregulatoryelementactivityinzebrafish
AT nefelidellepiane quantitativespatialandtemporalassessmentofregulatoryelementactivityinzebrafish
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