title: RNAi mediated silencing of ATPase RNA helicase gene in adult filarial parasite Brugia malayi impairs in vitro microfilaria release and adult parasite viability.
abstract: The DExD/H box families of RNA helicases are a multifunctional group of proteins involved in unwinding of inter- and intra-molecular base-paired regions. Successful knockdown of DEAD box RNA helicase gene (BmL3-Helicase) of human lymphatic filarial parasite Brugia malayi was done with specifically designed and chemically synthesized siRNA of <20bp to observe the role of enzyme in parasite biology and its worth as an antifilarial drug target. We made efforts to deliver siRNA into parasite by both electroporation and soaking that resulted into diminished helicase gene expression associated with decreased parasite motility, viability (97%) and release of microfilariae (81.0% reduction) from adult females in vitro. The specific gene knockdown also resulted into death of adult male worms in addition to phenotypic deformities in female worm intrauterine stages. RT-PCR of siRNA treated worms revealed a complete knockdown of BmL3-Helicase transcription within 16h. The present findings thus illustrate that targeting helicase gene of B. malayi would not only interfere with embryogenesis and microfilarial production but also result into decreased motility and viability of microfilariae and adult parasites. The B. malayi helicase enzyme thus represents a possible antifilarial drug target.
authors.affiliation: Division of Parasitology, Central Drug Research Institute, CSIR, M.G. Marg, Lucknow 226001 (U.P.), India.
authors.forename: Meghna
authors.initials: M
authors.lastname: Singh
authors.forename: Prashant Kumar
authors.initials: PK
authors.lastname: Singh
authors.forename: Shailja
authors.initials: S
authors.lastname: Misra-Bhattacharya
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D000704:Analysis of Variance; D000818:Animals; D017931:DNA Primers / Q000235:genetics; D018274:Electroporation; D005260:Female; D055785:Gene Knockdown Techniques; D008297:Male; D008842:Microfilariae / Q000235:genetics / Q000502:physiology*; D020365:RNA Helicases / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017174:Spirurida / Q000201:enzymology* / Q000502:physiology; D013778:Tetrazolium Salts; D013844:Thiazoles
chemical_list: D017931:DNA Primers; D034741:RNA, Small Interfering; D013778:Tetrazolium Salts; D013844:Thiazoles; D000251:Adenosine Triphosphatases; D020365:RNA Helicases; C022616:thiazolyl blue