Collins, C.R., Das, S., Wong, E.H., Andenmatten, N., Stallmach, R., Hackett, F., Herman, J.-P., Muller, S., Meissner, M. and Blackman, M.J. (2013) Robust inducible Cre recombinase activity in the human malaria parasite Plasmodium falciparumenables efficient gene deletion within a single asexual erythrocytic growth cycle. Molecular Microbiology, 88(4), pp. 687-701. (doi: 10.1111/mmi.12206)
|
Text
77976.pdf - Published Version 988kB |
Abstract
Asexual blood stages of the malaria parasite, which cause all the pathology associated with malaria, can readily be genetically modified by homologous recombination, enabling the functional study of parasite genes that are not essential in this part of the life cycle. However, no widely applicable method for conditional mutagenesis of essential asexual blood-stage malarial genes is available, hindering their functional analysis. We report the application of the DiCre conditional recombinase system to Plasmodium falciparum, the causative agent of the most dangerous form of malaria. We show that DiCre can be used to obtain rapid, highly regulated site-specific recombination in P. falciparum, capable of excising loxP-flanked sequences from a genomic locus with close to 100% efficiency within the time-span of a single erythrocytic growth cycle. DiCre-mediated deletion of the SERA5 3' UTR failed to reduce expression of the gene due to the existence of alternative cryptic polyadenylation sites within the modified locus. However, we successfully used the system to recycle the most widely used drug resistance marker for P. falciparum, human dihydrofolate reductase, in the process producing constitutively DiCre-expressing P. falciparum clones that have broad utility for the functional analysis of essential asexual blood-stage parasite genes.
Item Type: | Articles (Other) |
---|---|
Status: | Published |
Refereed: | Yes |
Glasgow Author(s) Enlighten ID: | Muller, Professor Sylke and Das, Dr Sujaan and Meissner, Professor Markus and Wong, Dr Eleanor |
Authors: | Collins, C.R., Das, S., Wong, E.H., Andenmatten, N., Stallmach, R., Hackett, F., Herman, J.-P., Muller, S., Meissner, M., and Blackman, M.J. |
College/School: | College of Medical Veterinary and Life Sciences > School of Infection & Immunity |
Journal Name: | Molecular Microbiology |
Journal Abbr.: | Mol. Microbiol. |
Publisher: | Wiley-Blackwell |
ISSN: | 0950-382X |
ISSN (Online): | 1365-2958 |
Published Online: | 26 March 2013 |
Copyright Holders: | Copyright © 2013 Blackwell Publishing |
First Published: | First published in Molecular Microbiology |
Publisher Policy: | Reproduced in accordance with the copyright policy of the publisher |
University Staff: Request a correction | Enlighten Editors: Update this record