PUBLICATION
Zebrafish nampt-a mutants are viable despite perturbed primitive hematopoiesis
- Authors
- Pomreinke, A.P., Müller, P.
- ID
- ZDB-PUB-240430-8
- Date
- 2024
- Source
- Hereditas 161: 1414 (Journal)
- Registered Authors
- Müller, Patrick
- Keywords
- none
- MeSH Terms
-
- Zebrafish*/genetics
- Nicotinamide Phosphoribosyltransferase*/genetics
- Mutation
- CRISPR-Cas Systems
- Animals
- Phenotype
- NAD/metabolism
- Zebrafish Proteins/genetics
- Gene Knockdown Techniques
- Morpholinos/genetics
- Hematopoiesis*/genetics
- PubMed
- 38685093 Full text @ Hereditas
Citation
Pomreinke, A.P., Müller, P. (2024) Zebrafish nampt-a mutants are viable despite perturbed primitive hematopoiesis. Hereditas. 161:1414.
Abstract
Background Nicotinamide phosphoribosyltransferase (Nampt) is required for recycling NAD+ in numerous cellular contexts. Morpholino-based knockdown of zebrafish nampt-a has been shown to cause abnormal development and defective hematopoiesis concomitant with decreased NAD+ levels. However, surprisingly, nampt-a mutant zebrafish were recently found to be viable, suggesting a discrepancy between the phenotypes in knockdown and knockout conditions. Here, we address this discrepancy by directly comparing loss-of-function approaches that result in identical defective transcripts in morphants and mutants.
Results Using CRISPR/Cas9-mediated mutagenesis, we generated nampt-a mutant lines that carry the same mis-spliced mRNA as nampt-a morphants. Despite reduced NAD+ levels and perturbed expression of specific blood markers, nampt-a mutants did not display obvious developmental defects and were found to be viable. In contrast, injection of nampt-a morpholinos into wild-type or mutant nampt-a embryos caused aberrant phenotypes. Moreover, nampt-a morpholinos caused additional reduction of blood-related markers in nampt-a mutants, suggesting that the defects observed in nampt-a morphants can be partially attributed to off-target effects of the morpholinos.
Conclusions Our findings show that zebrafish nampt-a mutants are viable despite reduced NAD+ levels and a perturbed hematopoietic gene expression program, indicating strong robustness of primitive hematopoiesis during early embryogenesis.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping