A genome-scale vector resource enables high-throughput reverse genetic screening in a malaria parasite

Gomes, A. R. et al. (2015) A genome-scale vector resource enables high-throughput reverse genetic screening in a malaria parasite. Cell Host and Microbe, 17(3), pp. 404-413. (doi: 10.1016/j.chom.2015.01.014) (PMID:25732065) (PMCID:PMC4362957)

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The genome-wide identification of gene functions in malaria parasites is hampered by a lack of reverse genetic screening methods. We present a large-scale resource of barcoded vectors with long homology arms for effective modification of the Plasmodium berghei genome. Cotransfecting dozens of vectors into the haploid blood stages creates complex pools of barcoded mutants, whose competitive fitness can be measured during infection of a single mouse using barcode sequencing (barseq). To validate the utility of this resource, we rescreen the P. berghei kinome, using published kinome screens for comparison. We find that several protein kinases function redundantly in asexual blood stages and confirm the targetability of kinases cdpk1, gsk3, tkl3, and PBANKA_082960 by genotyping cloned mutants. Thus, parallel phenotyping of barcoded mutants unlocks the power of reverse genetic screening for a malaria parasite and will enable the systematic identification of genes essential for in vivo parasite growth and transmission.

Item Type:Articles
Glasgow Author(s) Enlighten ID:Modrzynska, Dr Katarzyna
Authors: Gomes, A. R., Bushell, E., Schwach, F., Girling, G., Anar, B., Quail, M. A., Herd, C., Pfander, C., Modrzynska, K., Rayner, J. C., and Billker, O.
College/School:College of Medical Veterinary and Life Sciences > School of Infection & Immunity
Journal Name:Cell Host and Microbe
ISSN (Online):1934-6069
Published Online:26 February 2015
Copyright Holders:Copyright © 2015 The Authors
First Published:First published in Cell Host and Microbe 17(3):404-413
Publisher Policy:Reproduced under a Creative Commons License

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