Getting started with shRNA: Considerations & recommendations
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.
Learning Objectives:
- 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
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