Silencing of essential genes by RNA interference in Haemonchus contortus

• Published in: Parasitology Vol. 139; no. 5; pp. 613 - 629
• Main Authors: ZAWADZKI, J. L., KOTZE, A. C., FRITZ, J.-A., JOHNSON, N. M., HEMSWORTH, J. E., HINES, B. M., BEHM, C. A.
• Format: Journal Article
• Language: English
• Published: Cambridge, UK Cambridge University Press 01.04.2012
• Subjects: Animals; Caenorhabditis elegans; Caenorhabditis elegans - genetics; E coli; Escherichia coli; Gene Silencing; Gene Transfer Techniques; Genes, Essential - genetics; Genes, Helminth - genetics; Haemonchus - genetics; Haemonchus contortus; Larvae; Life Cycle Stages - genetics; Nematoda; Nematodes; RNA Interference; dsRNA; Haemonchus contortus; Caenorhabditis elegans; RNAi; RNA interference; gene knockdown; electroporation
• ISSN: 0031-1820; 1469-8161; 1469-8161
• DOI: 10.1017/S0031182012000121

In this study we assessed three technologies for silencing gene expression by RNA interference (RNAi) in the sheep parasitic nematode Haemonchus contortus. We chose as targets five genes that are essential in Caenorhabditis elegans (mitr-1, pat-12, vha-19, glf-1 and noah-1), orthologues of which are present and expressed in H. contortus, plus four genes previously tested by RNAi in H. contortus (ubiquitin, tubulin, paramyosin, tropomyosin). To introduce double-stranded RNA (dsRNA) into the nematodes we tested (1) feeding free-living stages of H. contortus with Escherichia coli that express dsRNA targetting the test genes; (2) electroporation of dsRNA into H. contortus eggs or larvae; and (3) soaking adult H. contortus in dsRNA. For each gene tested we observed reduced levels of mRNA in the treated nematodes, except for some electroporation conditions. We did not observe any phenotypic changes in the worms in the electroporation or dsRNA soaking experiments. The feeding method, however, elicited observable changes in the development and viability of larvae for five of the eight genes tested, including the ‘essential’ genes, Hc-pat-12, Hc-vha-19 and Hc-glf-1. We recommend the E. coli feeding method for RNAi in H. contortus and provide recommendations for future research directions for RNAi in this species.