PUBLICATION
Generation and Molecular Characterization of Transient tet1/2/3 Zebrafish Knockouts
- Authors
- Ross, S.E., Bogdanovic, O.
- ID
- ZDB-PUB-210520-8
- Date
- 2021
- Source
- Methods in molecular biology (Clifton, N.J.) 2272: 281-317 (Other)
- Registered Authors
- Bogdanovic, Ozren, Ross, Sam
- Keywords
- CRISPR/Cas9, DNA demethylation, DNA methylation, Embryogenesis, Reduced representation bisulfite sequencing (RRBS), TET proteins, Zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified/genetics
- Animals, Genetically Modified/growth & development
- Animals, Genetically Modified/metabolism*
- DNA Methylation*
- Dioxygenases/genetics
- Dioxygenases/metabolism*
- Gene Expression Regulation, Developmental*
- Zebrafish/genetics
- Zebrafish/growth & development
- Zebrafish/metabolism*
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism*
- PubMed
- 34009621 Full text @ Meth. Mol. Biol.
Citation
Ross, S.E., Bogdanovic, O. (2021) Generation and Molecular Characterization of Transient tet1/2/3 Zebrafish Knockouts. Methods in molecular biology (Clifton, N.J.). 2272:281-317.
Abstract
5-methylcytosine (5mC) is a gene-regulatory mark associated with transcriptional repression. 5mC can be erased through the catalytic action of Ten-eleven translocation (TET) methylcytosine dioxygenases (TET1, TET2, TET3), which oxidize 5mC resulting in its removal from the genome. In vertebrates, TET enzymes facilitate DNA demethylation of regulatory regions linked to genes involved in developmental processes. Consequently, TET ablation leads to severe morphological defects and developmental arrest. Here we describe a system that can facilitate the study of relationships between TET enzymes, 5mC, and embryo development. We provide detailed descriptions for the generation of F0 zebrafish tet1/2/3 knockouts using CRISPR/Cas9 technology and elaborate on the strategies to assess the impact of TET loss by reduced representation bisulfite sequencing (RRBS).
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping