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Related Experiment Video

Updated: Dec 15, 2025

Electroporation Method for In Vivo Delivery of Plasmid DNA in the Adult Zebrafish Telencephalon
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Efficiency and cell viability implications using tip type electroporation in zebrafish sperm cells.

Larissa O Daneluz1, Izani B Acosta2,3, Leandro S Nunes1

  • 1Laboratório de Genômica Estrutural, Programa de Pós-Graduação em Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.

Molecular Biology Reports
|July 15, 2020
PubMed
Summary

Tip type electroporation significantly improves DNA transfection efficiency in zebrafish sperm. This method enhances gene transfer with minimal negative impacts on sperm health and functionality.

Keywords:
Danio rerioElectroporationExogenous DNASMGTSpermatozoa

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Area of Science:

  • Aquaculture
  • Genetics
  • Reproductive Biology

Background:

  • Sperm-mediated gene transfer (SMGT) is a promising technique for zebrafish transgenesis.
  • Current methods for transfecting fish sperm require improvement for efficiency and safety.

Purpose of the Study:

  • To demonstrate the feasibility of tip type electroporation for zebrafish sperm transfection.
  • To establish a protocol yielding high transfection efficiency with minimal side effects.

Main Methods:

  • Zebrafish sperm were transfected with Cy3-labelled DNA using tip type electroporation (500–1500 V).
  • Sperm kinetics were analyzed via Computer Assisted Semen Analysis (CASA).
  • Cell integrity, reactive oxygen species (ROS), mitochondrial functionality, and transfection rates were assessed using flow cytometry.

Main Results:

  • Transfection rates increased with voltage, reaching 86.6% ± 1.9 at 1500 V.
  • Motility and plasma membrane integrity decreased with increasing field strength (P < 0.05).
  • ROS levels and mitochondrial functionality remained unaffected by electroporation (P > 0.05).

Conclusions:

  • Tip type electroporation effectively enhances exogenous DNA internalization in zebrafish sperm.
  • The protocol achieves high transfection rates with acceptable levels of sperm damage.