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Optimized transgenesis in Xenopus laevis/gilli isogenetic clones for immunological studies
Author(s) -
Nedelkovska Hristina,
Robert Jacques
Publication year - 2012
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/dvg.20809
Subject(s) - transgenesis , biology , xenopus , transgene , germline , genetics , transposase , gene , computational biology , genome , reproductive technology , transposable element , embryogenesis
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. genesis 50:300–306, 2012. © 2011 Wiley Periodicals, Inc.