UCL Cancer Institute
2 ARTICLES PUBLISHED IN JoVE
Biology
Generation of Stable Human Cell Lines with Tetracycline-inducible (Tet-on) shRNA or cDNA Expression
Marta Gomez-Martinez 1, Debora Schmitz 2, Alexander Hergovich 1
1UCL Cancer Institute, 2Friedrich Miescher Institute for Biomedical Research
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.
Biology
Workflow for High-content, Individual Cell Quantification of Fluorescent Markers from Universal Microscope Data, Supported by Open Source Software
Simon R. Stockwell 1, Sibylle Mittnacht 1
1Cancer Biology, UCL Cancer Institute
Presented is a flexible informatics workflow enabling multiplexed image-based analysis of fluorescently labeled cells. The workflow quantifies nuclear and cytoplasmic markers and computes marker translocation between these compartments. Procedures are provided for perturbation of cells using siRNA and reliable methodology for marker detection by indirect immunofluorescence in 96-well formats.
