DEVELOPMENT OF AN IN VITRO RNAI TO INVESTIGATE GENE FUNCTION IN BRUGIA MALAYI | ||||
Journal of the Egyptian Society of Parasitology | ||||
Article 26, Volume 46, Issue 3, December 2016, Page 717-728 PDF (538.04 K) | ||||
Document Type: Original Article | ||||
DOI: 10.21608/jesp.2016.88330 | ||||
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Authors | ||||
ADEL MOHAMMED FARGHALY1; HANY ABD EL-HAI AFIFY1; AFAF ABD EL-RAOUF TAHA1; EMAN MOSTAFA2, 3 | ||||
1Department of Parasitology, Faculty of Medicine, Zagazig University, Egypt. | ||||
2Department of Parasitology, Faculty of Medicine, Zagazig University, Egypt.+ | ||||
3Department 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 | ||||
Brugia malayi; Filarial nematode; Gene function; RNAi | ||||
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