Abstract
High-throughput genetic screens interfering with gene expression are invaluable tools to identify gene function and phenotype-to-genotype interactions. Implementing such screens in the laboratory is challenging, and the choice between currently available technologies based on RNAi and CRISPR/Cas9 (CRISPR-associated protein 9) is not trivial. Identifying reliable candidate hits requires a streamlined experimental setup adjusted to the specific biological question. Here, we provide a critical assessment of the various RNAi/CRISPR approaches to pooled screens and discuss their advantages and pitfalls. We specify a set of best practices for key parameters enabling a reproducible screen and provide a detailed overview of analysis methods and repositories for identifying the best candidate gene hits.
Original language | English |
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Pages (from-to) | 38-55 |
Number of pages | 18 |
Journal | Trends in Biotechnology |
Volume | 37 |
Issue number | 1 |
DOIs | |
Publication status | Published - Jan 2019 |
Keywords
- CRISPR/Cas9
- CRISPRa
- CRISPRi
- RNAi
- essential genes
- library pooled screens