PUBLICATION
Generation and validation of a myoglobin knockout zebrafish model
- Authors
- Hejlesen, R., Kjær-Sørensen, K., Fago, A., Oxvig, C.
- ID
- ZDB-PUB-231018-52
- Date
- 2023
- Source
- Transgenic Research 32(6): 537-546 (Journal)
- Registered Authors
- Keywords
- CRISPR/Cas, Genetic compensation, Knockout, Myoglobin, Zebrafish
- MeSH Terms
-
- Animals
- CRISPR-Cas Systems
- Gene Knockout Techniques
- Mice
- Myoglobin*/genetics
- Myoglobin*/metabolism
- Phenotype
- Zebrafish*/genetics
- Zebrafish*/metabolism
- Zebrafish Proteins/genetics
- PubMed
- 37847464 Full text @ Transgenic. Res.
Citation
Hejlesen, R., Kjær-Sørensen, K., Fago, A., Oxvig, C. (2023) Generation and validation of a myoglobin knockout zebrafish model. Transgenic Research. 32(6):537-546.
Abstract
Previous studies using myoglobin (Mb) knockout mice and knockdown zebrafish have presented conflicting results about in vivo phenotypes resulting from the loss of this conserved and highly expressed protein, and therefore a new well-characterized knockout model is warranted. We here describe the generation of three distinct zebrafish mb knockout lines using the CRISPR/Cas system. None of the three lines exhibited any morphological phenotypes, changes in length, or lethality during embryonic and larval development. The adult homozygous knockout mb(Auzf13.2) zebrafish line were absent of Mb protein, had an almost complete degradation of mb mRNA, and showed no changes in viability, length, or heart size. Furthermore, transcriptomic analysis of adult heart tissue showed that mb knockout did not cause altered expression of other genes. Lastly, no off-targeting was observed in 36 screened loci. In conclusion, we have generated three mb knockout lines with indistinguishable phenotypes during embryonic and larval development and validated one of these lines, mb(Auzf13.2), to have no signs of genetic compensation or off-target effects in the adult heart. These findings suggests that the mb(Auzf13.2) shows promise as a candidate for investigating the biological role of Mb in zebrafish.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping