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Kajumbulaet al. Genetic Vaccines and Therapy2012,10:8 http://www.gvtjournal.com/content/10/1/8
R E S E A R C HOpen Access Targeting wildtype Erythrocyte receptors for PlasmodiumfalciparumandvivaxMerozoitesby Zinc Finger NucleasesIn silico: Towards a Genetic Vaccine against Malaria 1 21,3* Henry Kajumbula , Wilson Byarugabaand Misaki Wayengera
Abstract Background:Malaria causes immense human morbidity and mortality globally. The plasmodium speciesvivaxand falciparumcause over 75 % clinical malaria cases. Until now, genebased strategies against malaria have only been applied to plasmodium species and their mosquitovector. Merozoites of these two respective plasmodiumspecies target and invade red blood cells (RBCs) by using the duffy antigen receptor for chemokines (DARC), and Sialic Acid (SLC4A1) residues of the Olinked glycans of Glycophorin A. RBCs of naturally selected duffynegative blacks are resistant to P.vivaxtropism. We hypothesized that artificial aberration of the hostpathway by target mutagenesis of either RBCreceptors, may abolish or reduce susceptibility of the host to malaria. As a first step towards the experimental actualization of these concepts, we aimed to identify zinc finger arrays (ZFAs) for constructing ZFNs that target genes of either wildtype hostRBC receptors. Methods:InSilicoGene & Genome Informatics Results:Using the genomic contextual nucleotidesequences of homosapiensdarc and glycophorina, and the ZFNconsortia software CoDAZiFiTZFA and CoDAZiFiTZFN: we identified 163 and over 1,000 single zinc finger arrays(sZFAs) that bind sequences within the genes for the two respective RBCreceptors. Second,2 and 18 paired zinc finger arrays (pZFAs) that are precursors for zinc finger nucleases (ZFNs) capable of cleaving the genes for darc and glycophorinawere respectively assembled.Third,a megaBLAST evaluation of the genomewide cleavage specificity of this set of ZFNs was done, revealing alternate homologous nucleotide targets in the human genome other than darc or glycophorin A. Conclusions:ZFNs engineered with these ZFAprecursorswith further optimization to enhance their specificity to only darc and glycophorina, could be used in constructing an experimental genebasedmalaria vaccine. Alternatively, meganucleases and transcription activatorlike (TAL) nucleases that target conserved stretches of darc and glycophorinaDNA may serve the purpose of abrogating invasion of RBCs byfalciparamandvivaxplasmodia species. Keywords:Malaria,Plasmodium, P.falciparum, P.vivax, Merozoites, RBCreceptors, Darc, Glycophorin A, Zinc finger nucleases, Hostpathway, Abrogation, Genetic vaccine
* Correspondence: wmisaki@yahoo.com 1 Dept of Medical Microbiology, School of Biomedical Science, College of Health Sciences, Makerere University, P O Box 7072, Kampala, Uganda 3 Unit of Genetics & Genomics, Dept of Pathology, School of Biomedical Science, College of Health Sciences, Makerere University, P O Box 7072, Kampala, Uganda Full list of author information is available at the end of the article
© 2012 Kajumbula et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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