Email updates

Keep up to date with the latest news and content from Genetic Vaccines and Therapy and BioMed Central.

Open Access Research

Targeting wild-type Erythrocyte receptors for Plasmodium falciparum and vivax Merozoites by Zinc Finger Nucleases In- silico: Towards a Genetic Vaccine against Malaria

Henry Kajumbula1, Wilson Byarugaba2 and Misaki Wayengera13*

Author Affiliations

1 Dept of Medical Microbiology, School of Biomedical Science, College of Health Sciences, Makerere University, P O Box 7072, Kampala, Uganda

2 Dept of Biochemistry, Kampala International University Western Campus, P O Box 61, Ishaka, Bushenyi, 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

For all author emails, please log on.

Genetic Vaccines and Therapy 2012, 10:8  doi:10.1186/1479-0556-10-8

Published: 31 August 2012

Abstract

Background

Malaria 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.

Methods

In-Silico Gene & Genome Informatics

Results

Using 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.

Conclusions

ZFNs 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.

Keywords:
Malaria; Plasmodium; P. falciparum; P. vivax; Merozoites; RBC-receptors; Darc; Glycophorin A; Zinc finger nucleases; Host-pathway; Abrogation; Genetic vaccine