Format

Send to

Choose Destination
Genesis. 2012 Mar;50(3):300-6. doi: 10.1002/dvg.20809. Epub 2011 Dec 27.

Optimized transgenesis in Xenopus laevis/gilli isogenetic clones for immunological studies.

Author information

1
Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, NY 14642, USA.

Abstract

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.

PMID:
21954010
PMCID:
PMC3250570
DOI:
10.1002/dvg.20809
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Wiley Icon for PubMed Central
Loading ...
Support Center