PUBLICATION

Transgenic expression in zebrafish embryos with an intact chorion by electroporation and microinjection

Authors
Tazin, N., Lambert, C.J., Samuel, R., Stevenson, T.J., Bonkowsky, J.L., Gale, B.K.
ID
ZDB-PUB-231017-57
Date
2023
Source
Biotechnology reports (Amsterdam, Netherlands)   40: e00814e00814 (Journal)
Registered Authors
Bonkowsky, Joshua
Keywords
DNA, Electroporation, Gene delivery, Microinjection, Zebrafish
MeSH Terms
none
PubMed
37840570 Full text @ Biotechnol Rep (Amst)
Abstract
Electroporation is regularly used to deliver agents into cells, including transgenic materials, but it is not used for mutating zebrafish embryos due to the lack of suitable systems, information on appropriate operating parameters, and the challenges posed by the protective chorion. Here, a novel method for gene delivery in zebrafish embryos was developed by combining microinjection into the space between the chorion and the embryo followed by electroporation. This method eliminates the need for chorion removal and injecting into the space between the chorion and embryo eliminates the need for finding and identifying key cell locations before performing an injection, making the process much simpler and more automatable. We also developed a microfluidic electroporation system and optimized electric pulse parameters for transgenesis of embryos. The study provided a novel method for gene delivery in zebrafish embryos that can be potentially implemented in a high throughput transgenesis or mutagenesis system.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping