Borisa Skrjabina (Minsteres Universitāte) lekcija 22.jūlijā

„New Technologies in transgenic mice generation” kopsavilkums.

The TRAnsgenic Mouse Facility (TRAM) in the Institute of Experimental Pathology (ZMBE), Unversity of Münster was established to produce transgenic and gene deleted (targeted) mouse models. The facility provides full range of services, starting from DNA bioinformatic analysis, cloning procedures, targeting vector design, ES cell manipulation, DNA and ES cells microinjection services to all WWU Münster researchers. Since it was established in 1996, TRAM has evolved from transgenic core unit to powerful full-service division providing comprehensive service, from gene accession number up to targeted heterozygous mouse generation.

All experimental work in producing mouse models, the unit performs itself. TRAM has generated more than 200 different transgenic mouse lines and more than 30 different gene targeted mouse lines. We are using the latest technologies. For example using the “Sleeping beauty” transposase system (Mates L. et al., 2009) for pronuclear injections increases efficiency of transgenic mice generation up to 50-70%. We are using the meganuclease mouse genome editing technology in order to increase efficiency of gene-targeted mouse model generation. For example, the ZFN approach (Perez-Pinera P. et al., 2011) dramatically increases efficiency of the ROSA26 locus targeting, and custom assembled TALEN nucleases facilitate efficiency in conventional gene targeting. We have modified the “Golden Gate” TALEN assembly destination vector (Cermak T. et al., 2011) and are using it efficiently for custom gene-targeted mouse models. Examples of using the “Sleeping Beauty”system or meganucleases in mouse genome modifications will be presented.