PUBLICATION
Quantitative spatial and temporal assessment of regulatory element activity in zebrafish
- Authors
- Bhatia, S., Kleinjan, D.J., Uttley, K., Mann, A., Dellepiane, N., Bickmore, W.A.
- ID
- ZDB-PUB-211120-5
- Date
- 2021
- Source
- eLIFE 10: (Journal)
- Registered Authors
- Bhatia, Shipra
- Keywords
- developmental biology, enhancer, gene regulation, genetics, genomics, human disease, live imaging, noncoding genome, transgenic assay, zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified/embryology
- Animals, Genetically Modified/genetics
- Embryo, Nonmammalian/metabolism
- Embryonic Development/genetics
- Regulatory Elements, Transcriptional*
- Zebrafish/embryology
- Zebrafish/genetics*
- PubMed
- 34796872 Full text @ Elife
Citation
Bhatia, S., Kleinjan, D.J., Uttley, K., Mann, A., Dellepiane, N., Bickmore, W.A. (2021) Quantitative spatial and temporal assessment of regulatory element activity in zebrafish. eLIFE. 10:.
Abstract
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping