|Production of new transgenic Hydra via embryo microinjection:|
The introduction of foreign DNA (transgenes) into the genome of Hydra allows studies of gene expression and protein function within live polyps. The Transgenic Hydra Facility at Kiel University offers the opportunity to produce such transgenic polyps.
The Facility will perform all the steps necessary to produce transgenic polyps, including set-up, embryo harvesting, and DNA microinjection. The investigator is responsible for supplying the purified construct (DNA). We will microinject minimum of 20 viable embryos. Once founder polyps have been identified, the Facility will ship the animals to the investigator. The Facility does not provide the mass culturing of transgenic polyps.
Essentially the steps necessary for the production of transgenic polyps are:
- Construction of a transgenic vector containing a suitable promoter, a genomic clone or DNA and intron fragment, and polyadenylation addition sequences
- Purification of the transgene for microinjection
- Microinjection of the transgene into the fertilized egg to generate polyps containing copies of that transgene integrated randomly in their genome.
Step 1 and 2 must be performed by the investigator. Step 3 will be done at the Facility in Kiel.
Purification of Transgene for Microinjection:
Highly purified DNA is essential for successful transgenic production. Particles within the DNA could easily clog the microinjection needle and/or interfere with integration of the DNA. We recommend that you isolate the fragment by using the Quiagen Midi Prep Kit and resuspend it in water.
The investigator will need to contact the scientific supervisor of the Facility. This consultation will enable us to determine the specific needs of the investigator, review the design of the transgenic vector, and discuss conditions for using this service. Please contact Alexander Klimovich, scientific coordinator.