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...
Guardado en:
Autores principales: | , , , , , |
---|---|
Formato: | article |
Lenguaje: | EN |
Publicado: |
eLife Sciences Publications Ltd
2021
|
Materias: | |
Acceso en línea: | https://doaj.org/article/cabea42dcfad455facb02d1a512d8384 |
Etiquetas: |
Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
|
id |
oai:doaj.org-article:cabea42dcfad455facb02d1a512d8384 |
---|---|
record_format |
dspace |
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 |
work_keys_str_mv |
AT shiprabhatia quantitativespatialandtemporalassessmentofregulatoryelementactivityinzebrafish AT dirkjankleinjan quantitativespatialandtemporalassessmentofregulatoryelementactivityinzebrafish AT kirstyuttley quantitativespatialandtemporalassessmentofregulatoryelementactivityinzebrafish AT anitamann quantitativespatialandtemporalassessmentofregulatoryelementactivityinzebrafish AT nefelidellepiane quantitativespatialandtemporalassessmentofregulatoryelementactivityinzebrafish AT wendyabickmore quantitativespatialandtemporalassessmentofregulatoryelementactivityinzebrafish |
_version_ |
1718420157660921856 |