DEVELOPMENT OF AN IN VITRO RNAI TO INVESTIGATE GENE FUNCTION IN BRUGIA MALAYI

Document Type : Original Article

Authors

1 Department of Parasitology, Faculty of Medicine, Zagazig University, Egypt.

2 Department of Parasitology, Faculty of Medicine, Zagazig University, Egypt.+

3 Department of Infectious Diseases2, University of Georgia, 501 D.W. Brooks Drive, Athens, GA, USA

Abstract

The RNA interference (RNAi) has the ability to turn off individual gene expression. So, it affords a remarkably specific tool for studying the effects of genes. It is regarded as a direct approach for determining such gene/genes functions and offers a valuable tool for modern drug discovery. The study aimed to develop in vitro RNAi in Brugia malayi with particular interest to study the function of Brugia malayi avr-14 (Bm-avr-14) and Brugia malayi β-tubulin (Bm-β-tubulin) genes. Bm-avr-14 is a gene encoding glutamate gated chloride channel (GluCl) which binds ivermectin and Bm-β-tubulin is a gene encoding β-tubulin which binds albendazole. Adult female worms were soaked in heterogeneous
sh rt interfering RNA (hsiRNA) with interest to study the role of two genes Bm-avr-14 and Bm-β- tubulin. Then, we assessed the knock down effects of target genes using worminator system and real time PCR. We found that worms treated with hsiRNA of Bm-avr-14 had a significant reduction in microfilariae (mf) production in comparison with untreated worms or worms treated with hsiRNA of green fluorescent protein (GFP). No significant reduction in mf production with Bm-β-tubulin gene was obtained. There were no changes in the movement of adults treated with either Bm-avr-14 or Bm-β-tubulin hsiRNAs. Inconsistent RNAi mediated suppression was achieved with Bm-avr-14 and Bm-β-
tubulin using real time PCR. The data may be helpful in assessment of drug target potential of genes.

Keywords

Journal of the Egyptian Society of Parasitology
Volume 46, Issue 3
December 2016
Pages 717-728

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