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Genetic manipulation of betta fish.

Alec Palmiotti1,2, Madison R Lichak1,2, Pei-Yin Shih1,2

  • 1Zuckerman Mind Brain Behavior Institute, Columbia University, New York, NY, United States.

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Researchers have developed new genetic tools for studying Siamese fighting fish (betta). These methods, including CRISPR and Tol2 transgenesis, enable gene knockout and knockin, advancing betta research.

Keywords:
CRISPRTol2 systembetta fishgenetic manipulationgenome editingknock inknock outtransgenesis methodologies

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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.