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

Abstract

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.

Keywords: Cancer; Cell Biology; Chromatin immunoprecipitation (ChIP); Gene Expression; Molecular Biology; Protein Biochemistry.