Optimized transgenesis in Xenopus laevis/gilli isogenetic clones for immunological studies.
Xenopus laevis provides a unique animal model, alternative to mouse, to study immunology. Even though, several methodologies have been developed for the generation of transgenic Xenopus, to date none have been adapted for the X. laevis/gilli (LG) isogenetic clones that are essential for immunological studies. Since LG clones are generated via gynogenesis, transgenic methods using transgene integration into the sperm nuclei are not suited. Therefore, we have tested three alternative methods for LG transgenesis: the phiC31 integrase, the Sleeping Beauty transposase, and the I-SceI meganuclease. All three techniques produced transgenic LG clones; however, the I-SceI meganuclease was most effective. It resulted in high transgenesis efficiency (35-50%), bright nonmosaic GFP expression as well as stable germline transmission with 100% of the progeny carrying the transgene. Production of transgenic LG clones will allow us to modulate immune gene expression and further strengthen X. laevis as a biomedical model.
PubMed ID: 21954010
PMC ID: PMC3250570
Article link: Genesis.
Grant support: 1R03-HD061671-01 NICHD NIH HHS , R24-AI-059830-06 NIAID NIH HHS , T32-AI 07285 NIAID NIH HHS , R03 HD061671-02 NICHD NIH HHS , R24 AI059830-06 NIAID NIH HHS , T32 AI007285-24 NIAID NIH HHS , R24 AI059830 NIAID NIH HHS , R03 HD061671 NICHD NIH HHS , T32 AI007285 NIAID NIH HHS
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