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RNAi/CRISPR Screens: from a Pool to a Valid Hit

Author:
Schuster, Anne, Erasimus, Hélène, Fritah, Sabrina, Nazarov, Petr V., van Dyck, Eric, Niclou, Simone P., Golebiewska, Anna
Source:
Trends in biotechnology 2019 v.37 no.1 pp. 38-55
ISSN:
0167-7799
Subject:
RNA interference, gene editing, gene expression, genes
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.
Agid:
6131740