A single vector containing modified cre recombinase and LOX recombination sequences for inducible tissue-specific amplification of gene expression.
The selective alteration of the genome using Cre recombinase to target the rearrangement of genes flanked by LOX recognition sequences has required the use of two separate genetic constructs in trans, one containing cre and the other containing the gene of interest flanked by LOX sites. We have developed a strategy in which both the cre recombinase gene and LOX recombination sites may be cloned within a single vector in cis. This method uses a modified form of Cre (CREM) that contains alterations to the 5' region including the introduction of a Kozak consensus sequence and insertion of a functional intron. This system allows for the inducible, tissue-specific activation or inactivation of gene expression in a single vector and can be utilized for the 300-fold amplification of gene expression from a weak promoter. This approach can be applied to targeting strategies for generating genetically altered mice and gene therapy.
Kaczmarczyk
S J
SJ
Transgenic Oncogenesis Group, Laboratory of Cell Regulation and Carcinogenesis, National Cancer Institute, Bethesda, MD 20892, USA.
Green
J E
JE
D000818:Animals; D001287:Attachment Sites, Microbiological / Q000235:genetics*; D017101:Bacteriophage P1 / Q000201:enzymology; D016384:Consensus Sequence / Q000235:genetics; D003587:Cytomegalovirus / Q000235:genetics; D005057:Eukaryotic Cells / Q000378:metabolism; D018965:Frameshifting, Ribosomal / Q000235:genetics; D005786:Gene Expression Regulation; D018390:Gene Targeting / Q000379:methods*; D017930:Genes, Reporter / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics*; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D007438:Introns / Q000235:genetics; D051379:Mice; D016254:Mutagenesis, Insertional / Q000235:genetics; D009928:Organ Specificity; D011387:Prokaryotic Cells / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D011995:Recombination, Genetic / Q000235:genetics*; D014158:Transcription, Genetic / Q000235:genetics; D014764:Viral Proteins
D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases