Application of an F0-based genetic assay in adult zebrafish to identify modifier genes of an inherited cardiomyopathy

Modifier genes contribute significantly to our understanding of pathophysiology in human diseases; however, effective approaches to identify modifier genes are still lacking. Here, we aim to develop a rapid F0-based genetic assay in adult zebrafish using the bag3 gene knockout (bag3e2/e2) cardiomyop...

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Bibliographic Details
Published in:Disease models & mechanisms 2023-05, Vol.16 (5)
Main Authors: Ding, Yonghe, Wang, Mingmin, Bu, Haisong, Li, Jiarong, Lin, Xueying, Xu, Xiaolei
Format: Article
Language:English
Subjects:
Adaptor Proteins, Signal Transducing - metabolism
Animals
Apoptosis Regulatory Proteins - genetics
bag3
Cardiomyopathies - genetics
cardiomyopathy
Gene Knockout Techniques
Genes, Modifier
microhomology-mediated end joining
modifier gene
Resource
RNA, Guide, CRISPR-Cas Systems
zebrafish
Zebrafish - genetics
Zebrafish - metabolism
Zebrafish Proteins - genetics
Zebrafish Proteins - metabolism
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Summary:Modifier genes contribute significantly to our understanding of pathophysiology in human diseases; however, effective approaches to identify modifier genes are still 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, by 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 inherited cardiomyopathy.
ISSN:1754-8403
1754-8411
DOI:10.1242/dmm.049427