Genetic manipulation of betta fish.

Area of Science:

• Aquatic biology
• Genetics
• Animal models

Background:

• Betta fish (Betta splendens) exhibit significant morphological diversity and aggressive behavior.
• Limited genomic manipulation tools have previously hindered functional studies in betta.
• Understanding betta genetics is crucial for research into their unique traits.

Purpose of the Study:

• To optimize and present three key genetic manipulation technologies for Betta splendens.
• To establish CRISPR/Cas9-mediated knockout and knockin capabilities.
• To implement Tol2-mediated transgenesis for gene expression studies.

Main Methods:

• CRISPR/Cas9-mediated gene knockout of alkal2l, bco1l, and mitfa.
• CRISPR/Cas9-mediated knockin of a fluorescent protein into the mitfa locus.
• Tol2-mediated transgenesis for ubiquitous GFP expression and heart-specific fluorescent protein expression.

Main Results:

• Successful knockout of alkal2l, bco1l, and mitfa, with analysis of their effects on betta viability and pigmentation.
• Demonstration of CRISPR/Cas9 knockin efficiency by inserting a fluorescent protein into the mitfa gene.
• Generation of transgenic betta lines with both ubiquitous and tissue-specific fluorescent protein expression.

Conclusions:

• The study provides optimized genetic tools for Betta splendens research.
• These methods facilitate functional genomic studies and the investigation of betta traits.
• The developed resources will accelerate research utilizing betta as an animal model.