Enhancer Detection

Thomas Becker, Sars International Centre, Bergen Norway | local teacher
Staale Ellingsen, EMBL Heidelberg, Germany | external teacher
Mary Laplante, Hiroshi Kikuta | local teaching assistants

tom.becker@sars.no | ellingse@embl.de

The generation of transgenic lines of fish expressing reporter genes encoding fluorescent proteins driven by specific promoters has been labor intensive, often taking a year or more per line. Using retroviral vectors it is now possible to generate hundreds to thousands of transgenic lines of fish in a relatively short time, allowing the collection of transgenic lines expressing the reporter in specific tissues under the control of enhancer elements of known as well as novel genes. Laboratory work in this part of the course includes production and microinjection of retroviral particles into zebrafish embryos, fluorescence microscopy screening and isolation of transgenic embryos with novel expression patterns, and isolation of flanking sequence from individual F1 embryos. This topic also includes live fluorescence imaging and data collection and data base management of hundreds of transgenic lines of fish in a single fish facility.

Suggested Readings
  • Ellingsen, S, Laplante, ML, Koenig, M, Kikuta, H, Furmanek, T,. Hoivik, E., and Becker, TS. Large-scale in vivo detection of cis-regulatory regions in the zebrafish genome by insertion of murine retroviral vectors. Submitted.