Use of an internal ribosome entry site for bicistronic expression of Cre recombinase or rtTA transactivator.
Conditional gene targeting depends on tissue and time specificity of recombination events. Endogenous promoters are often used to drive various transgenic constructs. To avoid the problems associated with reconstituting a specific expression pattern in transgenic animals by this method, we tested the internal ribosome entry site of the encephalomyocarditis virus, to enable linkage of the Cre recombinase or rtTA trans-activator to 3' untranslated ends of endogenous genes. Here we report that these constructs function effectively in COS cells. The data suggest that these cassettes will be appropriate for 3' targeting of mouse genes.
Department of Neuroscience, Uppsala University, Biomedical Center, Box 587, S-751 23 Uppsala, Sweden. jonas.lindeberg@mun.uu.se
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Ebendal
D000818:Animals; D001426:Bacterial Proteins; D019556:COS Cells; D004686:Encephalomyocarditis virus / Q000235:genetics; D015870:Gene Expression; D018390:Gene Targeting; D017405:Herpes Simplex Virus Protein Vmw65 / Q000096:biosynthesis / Q000235:genetics; D019426:Integrases / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D014176:Protein Biosynthesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012270:Ribosomes / Q000378:metabolism*; D015534:Trans-Activators / Q000096:biosynthesis* / Q000235:genetics; D014764:Viral Proteins
D001426:Bacterial Proteins; D017405:Herpes Simplex Virus Protein Vmw65; D011993:Recombinant Fusion Proteins; C116102:TetR protein, Clostridium tetani; D015534:Trans-Activators; D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases