Abstract
BackgroundMalaria causes immense human morbidity and mortality globally. The plasmodium species vivax and falciparum cause 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 plasmodium species 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.vivax tropism. We hypothesized that artificial aberration of the host-pathway by target mutagenesis of either RBC –receptors, 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.MethodsIn-Silico Gene & Genome InformaticsResultsUsing the genomic contextual nucleotide-sequences of homo-sapiens darc 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-a were 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.ConclusionsZFNs engineered with these ZFA-precursors--with 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-a DNA may serve the purpose of abrogating invasion of RBCs by falciparam and vivax plasmodia species.
Highlights
Malaria causes immense human morbidity and mortality globally
Identity of unpaired zinc finger arrays to bind darc and glycophorin-a sequences We identified 165 and close to 1,000 unpaired or single zinc finger arrays that bind the sequential nucleotide sequences constituting the genes for the two erythrocyte receptors for the merozoites of P. vivax and - falciparum
We explore zinc finger arrays (ZFAs)-precursors of zinc finger nucleases (ZFNs) that may be used for target mutagenesis and abrogation of red blood cells (RBCs)-receptors for merozoites of two major malaria causing plasmodium
Summary
Malaria causes immense human morbidity and mortality globally. The plasmodium species vivax and falciparum cause over 75 % clinical malaria cases. Gene-based strategies against malaria have only been applied to plasmodium species and their mosquito-vector. Merozoites of these two respective plasmodium species 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. Malaria is an infectious cause of immense humanmorbidity and mortality world-over [1]. Despite presence of effective Artemisinin-based combination chemotherapy for treating clinical malaria, the disease still claims over 1 million lives annually, most-children under the ages of 5 years [2,3]. Within malaria endemic regions of the tropics and sub-tropics, clinical malaria is responsible for up to: 40 % of public health expenditures, 30 % to 50 % of inpatient hospital admissions, and 60 % of outpatient health clinic visits [1,2,3,4]
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