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Engineering an fgfr4 knockout zebrafish to study its role in development and disease
Emma N Harrison1, Amanda N Jay1,2, Matthew R Kent1
1Center for Childhood Cancer, Nationwide Children's Hospital, Columbus, OH, United States of America.
Plos One
|November 22, 2024
Summary
Fibroblast growth factor receptor 4 (FGFR4) knockout zebrafish were generated. Homozygous mutants were smaller than wildtype fish, indicating FGFR4’s role in vertebrate growth.
Area of Science:
- Genetics and Developmental Biology
- Cancer Research
- Molecular Biology
Background:
- Fibroblast growth factor receptor 4 (FGFR4) plays a role in lipid metabolism, tissue repair, and vertebrate development.
- FGFR4 overexpression and mutations are linked to adult and pediatric cancers, making it a therapeutic target.
Purpose of the Study:
- To generate and characterize fgfr4 knockout zebrafish strains using CRISPR/Cas9.
- To investigate the role of FGFR4 signaling in vertebrate development and its potential as a cancer therapeutic target.
Main Methods:
- CRISPR/Cas9 gene editing was used to create three fgfr4 knockout zebrafish alleles.
- Single-cell wildtype zebrafish embryos were injected with fgfr4 targeting guide RNA and Cas9 proteins.
- Mutant strains were confirmed by assessing fgfr4 mRNA expression and resulting protein truncation.
Main Results:
- Three fgfr4 knockout zebrafish strains were successfully generated, with mutations leading to truncated FGFR4 proteins.
- All mutant strains showed significantly decreased fgfr4 mRNA expression.
- Homozygous fgfr4 mutant zebrafish were significantly smaller than wildtype fish at three months post fertilization.
Conclusions:
- The generated fgfr4 knockout zebrafish lines are valuable tools for studying FGFR4's role in development.
- These zebrafish models can aid in evaluating FGFR4 as a therapeutic target for cancers and other diseases.
