Control of siRNA expression utilizing Cre-loxP recombination system | Zendy

Vivi Kasim | Zendy; Makoto Miyagishi | Zendy; Kazunari Taira | Zendy

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Control of siRNA expression utilizing Cre-loxP recombination system

Author(s) -

Vivi Kasim,

Makoto Miyagishi,

Kazunari Taira

Publication year - 2003

Publication title -

nucleic acids symposium series

Language(s) - English

Resource type - Journals

eISSN - 1746-8272

pISSN - 0261-3166

DOI - 10.1093/nass/3.1.255

Subject(s) - cre recombinase , luciferase , rna interference , microbiology and biotechnology , nuclear localization sequence , small interfering rna , transcription (linguistics) , promoter , cre lox recombination , biology , expression vector , gene silencing , nls , rna , gene expression , transfection , genetics , cell culture , gene , transgene , recombinant dna , linguistics , philosophy , genetically modified mouse , cytoplasm

Vector-mediated systems for specific siRNA expression in mammalian cells using pol III promoters allowing high level of transcription activity have been developed in the past years, widening the usage of RNA interference (RNAi). In this study, we controlled the pol III promoter (U6 promoter)-driven expression of siRNA using the Cre-loxP system. Our "Cre-On" siRNA-expression vector against firefly luciferase activity could be switched on only in the presence of Cre recombinase, which, in this study, was delivered directly from the medium into the cells as TAT-NLS-Cre, a fusion protein with TAT peptide (an Arg rich peptide derived from HIV) and nuclear localizing signal (NLS). Upon the addition of TAT-NLS-Cre, complete and functional siRNAs were generated and reporter activity was suppressed.

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