PUBLICATION
Development of an endogenously myc-tagged TARDBP (TDP-43) zebrafish model using the CRISPR/Cas9 system and homology directed repair
- Authors
- Légaré, V.P., Rampal, C.J., Gurberg, T.J.N., Harji, Z.A., Allard-Chamard, X., Rodríguez, E.C., Armstrong, G.A.B.
- ID
- ZDB-PUB-220518-8
- Date
- 2022
- Source
- Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology 261: 110756 (Journal)
- Registered Authors
- Armstrong, Gary A.B.
- Keywords
- CRISPR, Homology directed repair, Motor neuron, TDP-43, Zebrafish
- MeSH Terms
-
- Animals
- CRISPR-Cas Systems
- DNA-Binding Proteins/genetics
- Epitopes/metabolism
- Neurodegenerative Diseases*/genetics
- Zebrafish*/genetics
- Zebrafish*/metabolism
- PubMed
- 35580804 Full text @ Comp. Biochem. Physiol. B Biochem. Mol. Biol.
Citation
Légaré, V.P., Rampal, C.J., Gurberg, T.J.N., Harji, Z.A., Allard-Chamard, X., Rodríguez, E.C., Armstrong, G.A.B. (2022) Development of an endogenously myc-tagged TARDBP (TDP-43) zebrafish model using the CRISPR/Cas9 system and homology directed repair. Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology. 261:110756.
Abstract
Many of the modern advances in cellular biology have been made by the expression of engineered constructs with epitope tags for subsequent biochemical investigations. While the utility of epitope tags has permitted insights in cellular and animal models, these are often expressed using traditional transgenic approaches. Using the CRISPR/Cas9 system and homology directed repair we recombine a single myc epitope sequence following the start codon of the zebrafish ortholog of TARDBP (TDP-43). TDP-43 is an RNA binding protein that is involved in the neurodegenerative disease amyotrophic lateral sclerosis and frontotemporal dementia. We report that zebrafish expressing the myc-tardbp engendered allele produced a stable protein that was detected by both western blot and immunofluorescence. Furthermore, both heterozygous and homozygous carriers of the myc-tardbp allele developed to sexual maturity. We propose that the methodology used here will be useful for zebrafish researchers and other comparative animal biologists interested in developing animal models expressing endogenously tagged proteins.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping