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Generation of Stable Human Cell Lines with Tetracycline-inducible (Tet-on) shRNA or cDNA Expression

Article DOI: 10.3791/50171-v • 09:51 min • March 5th, 2013
March 5th, 2013 •
1, 2, 1
1UCL Cancer Institute, 2Friedrich Miescher Institute for Biomedical Research

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Summary
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A rapid and simple way to generate human cell lines with inducible and reversible cDNA overexpression or shRNA-mediated knock-down of the gene of interest. This method enables researchers to reliably and highly reproducibly manipulate cell lines that are difficult to alter by transient transfection methods or conventional knockdown/knockout strategies.

Tags

Tetracycline-inducible Tet-on System ShRNA Expression CDNA Expression Stable Human Cell Lines Gene Manipulation Gene Expression Transient Overexpression Knockdown TetR Protein Regulatory Systems Transcription Initiation Tet-operator Promoter Region Tet-off System Tet-on System Tetracycline Presence Vector Systems RNA Interference (RNAi)
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