A method to inhibit the growth of Plasmodium falciparum by double-stranded RNA-mediated gene silencing of helicases.
Malaria in human is caused by four Plasmodium species, with Plasmodium falciparum responsible for the most severe form of the disease. Global resistance to multiple antimalarial drugs is becoming a major challenge in worldwide efforts to control malaria. It is essential to identify new targets. One possible target is helicases, which are important ubiquitous unwinding enzymes required for nucleic acid metabolism and the maintenance of genomic stability. Helicases are motor proteins that use the energy derived from their intrinsic nucleic acid-dependent NTPase activity to unwind the duplex nucleic acid substrate. In this chapter, we study the functional role of helicases in malaria parasite by using specific dsRNA against PfH45, one of the parasite helicases. We describe the methods for Plasmodium falciparum culture, the amplification of specific helicase gene, the construction of specific dsRNA, and the analysis of the effect of dsRNA on parasite growth. Using this approach, we show that helicases are indispensable enzymes, which are required for growth and most probably survival of the malaria parasite.
Malaria Group, International Centre for Genetic Engineering and Biotechnology, New Delhi, India.
Renu
R
Tuteja
D000818:Animals; D004265:DNA Helicases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D020868:Gene Silencing; D006801:Humans; D010963:Plasmodium falciparum / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D012330:RNA, Double-Stranded / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D012330:RNA, Double-Stranded; D011993:Recombinant Fusion Proteins; D004265:DNA Helicases