High Throughput siRNA Screening Using Reverse Transfection

Methods Mol Biol. 2016:1470:25-37. doi: 10.1007/978-1-4939-6337-9_3.

Abstract

RNA interference (RNAi) is a commonly used technique to knockdown gene function. Here, we describe a high throughput screening method for siRNA mediated gene silencing of the breast cancer cell line MDA-MB-231 using reverse transfection. Furthermore, we describe the setup for two separate methods for detecting viable and dead cells using either homogenous assays or image-based analysis.

Keywords: Acoustic droplet ejection; Gene silencing; High throughput screening; RNAi; Reverse transfection; siRNA screening.

MeSH terms

  • Cell Line, Tumor
  • Female
  • High-Throughput Screening Assays / methods*
  • Humans
  • RNA, Small Interfering* / genetics
  • Transfection / instrumentation
  • Transfection / methods*

Substances

  • RNA, Small Interfering