Short-hairpin RNAs (shRNAs) are routinely used in RNAi experiments to examine the specific roles of genes in biological processes. Combining shRNA with lentiviral vector-based delivery has become an invaluable tool for performing RNAi in difficult-to-transfect cells and in developing stable cell lines for long-term gene knock-down. In this presentation we discuss shRNA vector design considerations that ensure consistent and reliable shRNA expression. These include: selection of the optimal promoter and markers, delivery optimization, and an overview of the variables that can directly impact the success of shRNA-based knockdown. We will also discuss advantages of using shRNA tools in functional genomic studies and highlight best experimental practices.

Attendees will learn:

  • Vector design considerations for reliable shRNA expression
  • Strategies to ensure the shRNA functionality and specificity
  • shRNA delivery optimization and basic assay design considerations
  • Address commonly encountered questions and challenges