PUBLICATION
An F0-based genetic assay in adult zebrafish for discovering modifier genes of an inherited cardiomyopathy
- Authors
- Ding, Y., Wang, M., Bu, H., Li, J., Lin, X., Xu, X.
- ID
- ZDB-PUB-220429-14
- Date
- 2022
- Source
- Disease models & mechanisms 16(5): (Journal)
- Registered Authors
- Ding, Yonghe, Lin, Xueying, Xu, Xiaolei
- Keywords
- Cardiomyopathy, F0, Microhomology-mediated end joining, Modifier gene, Zebrafish, bag3
- MeSH Terms
-
- Adaptor Proteins, Signal Transducing/metabolism
- Animals
- Apoptosis Regulatory Proteins/genetics
- Cardiomyopathies*/genetics
- Gene Knockout Techniques
- Genes, Modifier
- RNA, Guide, Kinetoplastida
- Zebrafish*/genetics
- Zebrafish*/metabolism
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism
- PubMed
- 35481478 Full text @ Dis. Model. Mech.
Citation
Ding, Y., Wang, M., Bu, H., Li, J., Lin, X., Xu, X. (2022) An F0-based genetic assay in adult zebrafish for discovering modifier genes of an inherited cardiomyopathy. Disease models & mechanisms. 16(5):.
Abstract
Modifier genes contribute significantly to our understanding of pathophysiology in human diseases; however, effective approaches to identify modifier genes have been lacking. Here, we aim to develop a rapid F0-based genetic assay in adult zebrafish using the bag3 gene knockout (bag3e2/e2) cardiomyopathy model as a paradigm. First, utilizing a classic genetic breeding approach, we identified dnajb6b as a deleterious modifier gene for bag3 cardiomyopathy. Next, we established an F0-based genetic assay in adult zebrafish through injection of predicted microhomology-mediated end joining (MMEJ)-inducing single guide RNA/Cas9 protein complex. We showed that effective gene knockdown is maintained in F0 adult fish, enabling recapitulation of both salutary modifying effects of the mtor haploinsufficiency and deleterious modifying effects of the dnajb6b gene on bag3 cardiomyopathy. We finally deployed the F0-based genetic assay to screen differentially expressed genes in the bag3 cardiomyopathy model. As a result, myh9b was identified as a novel modifier gene for bag3 cardiomyopathy. Together, these data prove the feasibility of an F0 adult zebrafish-based genetic assay that can be effectively used to discover modifier genes for an inherited cardiomyopathy model.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping