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Modified gateway system for double shRNA expression and Cre/lox based gene expression by Nikolina Radulovich; Lisa Leung; Ming-Sound Tsao is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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## Background The growing need for functional studies of genes has set the stage for the development of versatile tools for genetic manipulations. ## Results Aiming to provide tools for high throughput analysis of gene functions, we have developed a modified short hairpin RNA (shRNA) and gene expression system based on Gateway Technology. The system contains a series of entry and destination vectors that enables easy transfer of shRNA or cDNA into lentiviral expression systems with a variety of selection or marker genes (i.e. puromycin, hygromycin, green fluorescent protein-EGFP, yellow fluorescent protein-YFP and red fluorescent protein-dsRed2). Our shRNA entry vector pENTR.hU6.hH1 containing two tandem human shRNA expression promoters, H1 and U6, was capable of co-expressing two shRNA sequences simultaneously. The entry vector for gene overexpression, pENTR.CMV.ON was constructed to contain CMV promoter with a multiple cloning site flanked by loxP sites allowing for subsequent Cre/lox recombination. Both shRNA and cDNA expression vectors also contained attL sites necessary for recombination with attR sites in our destination expression vectors. As proof of principle we demonstrat

Who reads Modified gateway system for double shRNA expression and Cre/lox based gene expression?

Author

Nikolina Radulovich; Lisa Leung; Ming-Sound Tsao

Publisher

BioMed Central; Springer (Biomed Central Ltd.); [London]: BioMed Central, 2001-; Springer Science and Business Media LLC (ISSN 1472-6750)

Published

2011

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)