Generation of stable Xenopus laevis transgenic lines expressing a transgene controlled by weak promoters.
Combining two existing protocols of trangenesis, namely the REMI and the I-SceI meganuclease methods, we generated Xenopus leavis expressing a transgene under the control of a promoter that presented a restricted pattern of activity and a low level of expression. This was realized by co-incubating sperm nuclei, the I-SceI enzyme and the transgene prior to transplantation into unfertilized eggs. The addition of the woodchuck hepatitis virus posttranscriptional regulatory element in our constructs further enhanced the expression of the transgene without affecting the tissue-specificity of the promoter activity. Using this combination of methods we produced high rates of fully transgenic animals that stably transmitted the transgene to the next generations with a transmission rate of 50% indicating a single integration event.
L'hostis-Guidet
Anne
A
Equipe SCANING, Interactions cellulaires et moléculaires, Rennes, France.
Recher
Gaëlle
G
Guillet
Brigitte
B
Al-Mohammad
Abdulrahim
A
Coumailleau
Pascal
P
Tiaho
François
F
Boujard
Daniel
D
Madigou
Thierry
T
D000818:Animals; D030801:Animals, Genetically Modified; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics; D017919:Hepatitis B Virus, Woodchuck / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000235:genetics; D011401:Promoter Regions, Genetic; D019076:Transgenes; D014982:Xenopus laevis / Q000235:genetics*
D051792:Basic Helix-Loop-Helix Proteins; D009419:Nerve Tissue Proteins; C093564:Neurogenic differentiation factor 1