Genome-scale RNAi screens for high-throughput phenotyping in bloodstream-form African trypanosomes : Nature Protocols : Nature Publishing Group

Genome-scale RNAi screens for high-throughput phenotyping in bloodstream-form African trypanosomes : Nature Protocols : Nature Publishing Group







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

• Lucy Glover,
• Sam Alsford,
• Nicola Baker,
• Daniel J Turner,
• Alejandro Sanchez-Flores,
• Sebastian Hutchinson,
• Christiane Hertz-Fowler,
• Matthew Berriman
• & David Horn

• Affiliations
• Contributions
• Corresponding author

Nature Protocols

 

10,

 

106–133

 

(2015)

 

doi:10.1038/nprot.2015.005

Published online

 

11 December 2014

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Abstract

• Abstract• 
• References• 
• Author information• 
• Supplementary information

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.

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