Optimized lentiviral vector transduction of adherent cells and analysis in sulforhodamine B proliferation and chromatin immunoprecipitation assays

Abstract

Summary: Transduction with lentiviral vectors is a useful approach to study the molecular function of specific genes in mammalian cells. Here, we present a calcium phosphate-based transfection protocol that guarantees highly efficient production and delivery of lentiviral vectors in adherent cultured cells. We also describe in detail a direct lysis technique to measure protein expression, an optimized sulforhodamine B proliferation assay, and a step-by-step chromatin immunoprecipitation procedure to verify the binding of ETV5 to E2F1 first intron in SYO-1 sarcoma cells.For complete details on the use and execution of this protocol, please refer to Kingston et al. (2003),1 Ireton et al. (2002),2 Brown et al. (2009),3 DeSalvo et al. (2021),4 Vichai and Kirtikara (2006),5 and Boyer et al. (2005).6 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cell Biology
• Cancer
• Molecular Biology
• Gene Expression
• Chromatin immunoprecipitation (ChIP)
• Protein Biochemistry