Send to

Choose Destination
Nat Protoc. 2015 Jan;10(1):106-33. doi: 10.1038/nprot.2015.005. Epub 2014 Dec 11.

Genome-scale RNAi screens for high-throughput phenotyping in bloodstream-form African trypanosomes.

Author information

Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee, Dundee, UK.
London School of Hygiene and Tropical Medicine, London, UK.
Oxford Nanopore Technologies, Oxford, UK.
Institute of Biotechnology, Universidad Nacional Autónoma de México, Cuernavaca, México.
Centre for Genomic Research, Institute of Integrative Biology, University of Liverpool, Liverpool, UK.
The Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK.


The ability to simultaneously assess every gene in a genome for a role in a particular process has obvious appeal. This protocol describes how to perform genome-scale RNAi library screens in bloodstream-form African trypanosomes, a family of parasites that causes lethal human and animal diseases and also serves as a model for studies on basic aspects of eukaryotic biology and evolution. We discuss strain assembly, screen design and implementation, the RNAi target sequencing approach and hit validation, and we provide a step-by-step protocol. A screen can yield from one to thousands of 'hits' associated with the phenotype of interest. The screening protocol itself takes 2 weeks or less to be completed, and high-throughput sequencing may also be completed within weeks. Pre- and post-screen strain assembly, validation and follow-up can take several months, depending on the type of screen and the number of hits analyzed.

[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Nature Publishing Group
Loading ...
Support Center