ZeGenesis – Generation of Genetic Models

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Tol2 Transgenic Models

Zebrafish transgenic lines can be generated by the random insertion of a desired DNA fragment, allowed by the activity of a Tol2 transposase. This is useful for expressing exogenous coding sequences (CDS) under the control of tissue-specific promoters. 

Technology used for generating:

  • Insertion of human genes´ CDSs
  • Reporter lines
  • Environmental sensors
  • Optogenetic tools

One-cell stage zebrafish embryos are injected with a mix containing tol2 mRNA and a plasmid containing the transgene of interest between two Tol2 sites.

During the early stages of embryonic development, the tol2 catalyzes the integration of the exogenous DNA into the genome of injected embryos.  If this process happens in cells that give rise to the germline, injected fish will be able to transmit the transgene to their progeny, thus allowing the establishment of a transgenic line.