Skip to main content
×
Home
    • Aa
    • Aa

Marker genes for activation of the RNA interference (RNAi) pathway in the free-living nematode Caenorhabditis elegans and RNAi development in the ovine nematode Teladorsagia circumcincta

  • T. Tzelos (a1) (a2), J.B. Matthews (a1), B. Whitelaw (a2) and D.P. Knox (a1)
Abstract
Abstract

The nematode Teladorsagiacircumcincta is a major cause of parasitic gastroenteritis in sheep in temperate regions. The development of resistance to the major anthelmintic classes used for its control is a threat to small ruminant farming sustainability. Vaccination is a potential alternative control method for this nematode. Gene datasets can be exploited to identify potential vaccine candidates and these validated further by methods such as RNA interference (RNAi) prior to vaccine trials. Previous reports indicate that RNAi in parasitic nematodes is inconsistent and, to date, there are no internal controls that indicate activation of the RNAi pathway in response to double-stranded RNA (dsRNA). The present aims were to determine whether or not the transcription levels of potential marker genes in the RNAi pathway could indicate activation of the pathway in Caenorhabditis elegans and to develop an RNAi platform in T. circumcincta. In C. elegans, transcript levels of three candidate marker genes, Ce-dcr-1 (Dicer), Ce-ego-1 (Enhancer of Glp-One family member) and Ce-rsd-3 (RNAi Spreading Defective), were analysed and results indicated that activation of the pathway had no effect on transcript levels of these genes. In T. circumcincta, two vaccine candidate genes from the Activation-associated Secreted Protein (ASP) family were targets for knockdown. RNAi experiments showed successful silencing of both targets, although inconsistencies in efficacy were observed. After testing a number of parameters that might affect variability, it was found that the length of the storage period of the larvae plays an important role in the consistency of the RNAi results.

Copyright
Corresponding author
*E-mail: Dave.Knox@moredun.ac.uk
Linked references
Hide All

This list contains references from the content that can be linked to their source. For a full set of references and notes please see the PDF or HTML where available.

E. Bernstein , A.A. Caudy , S.M. Hammond & G.J. Hannon (2001) Role for a bidentate ribonuclease in the initiation step of RNA interference. Nature 409, 363366.

J.J. Dalzell , P. McVeigh , N.D. Warnock , M. Mitreva , D.M. Bird , P. Abad , C.C. Fleming , T.A. Day , A. Mousley , N.J. Marks & A.G. Maule (2011) RNAi effector diversity in nematodes. PLoS Neglected Tropical Diseases 5, e1176.

P. Geldhof , E. Claerebout , D. Knox , I. Vercauteren , A. Looszova & J. Vercruysse (2002) Vaccination of calves against Ostertagia ostertagi with cysteine proteinase enriched protein fractions. Parasite Immunology 24, 263270.

P. Geldhof , I. Vercauteren , K. Gevaert , A. Staes , D.P. Knox , J. Vandekerckhove , J. Vercruysse & E. Claerebout (2003) Activation-associated secreted proteins are the most abundant antigens in a host protective fraction from Ostertagia ostertagi. Molecular and Biochemical Parasitology 128, 111114.

P. Geldhof , C. Whitton , W.F. Gregory , M. Blaxter & D.P. Knox (2005) Characterisation of the two most abundant genes in the Haemonchus contortus expressed sequence tag dataset. International Journal for Parasitology 35, 513522.

P. Geldhof , L. Murray , A. Couthier , J.S. Gilleard , G. McLauchlan , D.P. Knox & C. Britton (2006b) Testing the efficacy of RNA interference in Haemonchus contortus. International Journal for Parasitology 36, 801810.

K. Ghosh & P.J. Hotez (1999) Antibody-dependent reductions in mouse hookworm burden after vaccination with Ancylostoma caninum secreted protein 1. Journal of Infectious Diseases 180, 16741681.

S.M. Hammond (2005) Dicing and slicing: The core machinery of the RNA interference pathway. FEBS Letters 579, 58225829.

J.M. Hawdon , S. Narasimhan & P.J. Hotez (1999) Ancylostoma secreted protein 2: cloning and characterization of a second member of a family of nematode secreted proteins from Ancylostoma caninum. Molecular and Biochemical Parasitology 99, 149165.

D. Hoogewijs , K. Houthoofd , F. Matthijssens , J. Vandesompele & J.R. Vanfleteren (2008) Selection and validation of a set of reliable reference genes for quantitative sod gene expression analysis in C. elegans. BMC Molecular Biology 9, .

A.S. Hussein , K. Kichenin & M.E. Selkirk (2002) Suppression of secreted acetylcholinesterase expression in Nippostrongylus brasiliensis by RNA interference. Molecular and Biochemical Parasitology 122, 9194.

Z. Issa , W.N. Grant , S. Stasiuk & C.B. Shoemaker (2005) Development of methods for RNA interference in the sheep gastrointestinal parasite, Trichostrongylus colubriformis. International Journal for Parasitology 35, 935940.

R.S. Kamath , A.G. Fraser , Y. Dong , G. Poulin , R. Durbin , M. Gotta , A. Kanapin , N. Le Bot , S. Moreno , M. Sohrmann , D.P. Welchman , P. Zipperlen & J. Ahringer (2003) Systematic functional analysis of the Caenorhabditis elegans genome using RNAi. Nature 421, 231237.

R. Laing , T. Kikuchi , A. Martinelli , I. Tsai , R. Beech , E. Redman , N. Holroyd , D. Bartley , H. Beasley , C. Britton , D. Curran , E. Devaney , A. Gilabert , M. Hunt , F. Jackson , S. Johnston , I. Kryukov , K. Li , A. Morrison , A. Reid , N. Sargison , G. Saunders , J. Wasmuth , A. Wolstenholme , M. Berriman , J. Gilleard & J. Cotton (2013) The genome and transcriptome of Haemonchus contortus, a key model parasite for drug and vaccine discovery. Genome Biology 14, R88.

I. Maeda , Y. Kohara , M. Yamamoto & A. Sugimoto (2001) Large-scale analysis of gene function in Caenorhabditis elegans by high-throughput RNAi. Current Biology 11, 171176.

J.M. Maniar & A.Z. Fire (2011) EGO-1, a C. elegans RdRP, modulates gene expression via production of mRNA-templated short antisense RNAs. Current Biology 21, 449459.

A.G. Maule , P. McVeigh , J.J. Dalzell , L. Atkinson , A. Mousley & N.J. Marks (2011) An eye on RNAi in nematode parasites. Trends in Parasitology 27, 505513.

R. Menon , R.B. Gasser , M. Mitreva & S. Ranganathan (2012) An analysis of the transcriptome of Teladorsagia circumcincta: its biological and biotechnological implications. Eleventh International Conference on Bioinformatics (InCoB2012): Computational Biology. BMC Genomics 13 (Suppl. 7), S10, doi:10.1186/1471-2164-13-S7-S10

A.J. Nisbet , D.P. Knox , C.M. McNair , L.I. Meikle , S.K. Smith , L.A. Wildblood & J.B. Matthews (2009) Immune recognition of the surface associated antigen, Tc-SAA-1, from infective larvae of Teladorsagia circumcincta. Parasite Immunology 31, 3240.

A.J. Nisbet , N.E.V. Bell , T.N. McNeilly , D.P. Knox , R.M. Maizels , L.I. Meikle , L.A. Wildblood & J.B. Matthews (2010a) A macrophage migration inhibitory factor-like tautomerase from Teladorsagia circumcincta (Nematoda: Strongylida). Parasite Immunology 32, 503511.

A.J. Nisbet , S.K. Smith , S. Armstrong , L.I. Meikle , L.A. Wildblood , R.J. Beynon & J.B. Matthews (2010b) Teladorsagia circumcincta: Activation-associated secreted proteins in excretory/secretory products of fourth stage larvae are targets of early IgA responses in infected sheep. Experimental Parasitology 125, 329337.

A.J. Nisbet , D.S. Zarlenga , D.P. Knox , L.I. Meikle , L.A. Wildblood & J.B. Matthews (2011) A calcium-activated apyrase from Teladorsagia circumcincta: an excretory/secretory antigen capable of modulating host immune responses? Parasite Immunology 33, 236243.

B. Samarasinghe , D.P. Knox & C. Britton (2011) Factors affecting susceptibility to RNA interference in Haemonchus contortus and in vivo silencing of an H11 aminopeptidase gene. International Journal for Parasitology 41, 5159.

N.D. Sargison (2011) Pharmaceutical control of endoparasitic helminth infections in sheep. Veterinary Clinics of North America. Food Animal Practice 27, 139156.

H.D.F.H. Schallig & M.A.W. Van Leeuwen (1997) Protective immunity to the blood-feeding nematode Haemonchus contortus induced by vaccination with parasite low molecular weight antigens. Parasitology 114, 293299.

W. Tawe , E. Pearlman , T.R. Unnasch & S. Lustigman (2000) Angiogenic activity of Onchocerca volvulus recombinant proteins similar to vespid venom antigen 5. Molecular and Biochemical Parasitology 109, 9199.

M. Tijsterman , R.C. May , F. Simmer , K.L. Okihara & R.H.A. Plasterk (2004) Genes required for systemic RNA interference in Caenorhabditis elegans. Current Biology 14, 111116.

V. Vance & H. Vaucheret (2001) RNA silencing in plants - defense and counterdefense. Science 292, 22772280.

J. Wang & S.K. Kim (2003) Global analysis of dauer gene expression in Caenorhabditis elegans. Development 130, 16211634.

J. Wrigley , M. McArthur , P.B. McKenna & B. Mariadass (2006) Resistance to a triple combination of broad-spectrum anthelmintics in naturally-acquired Ostertagia circumcincta infections in sheep. New Zealand Veterinary Journal 54, 4749.

B. Zhan , Y. Liu , M. Badamchian , A. Williamson , J. Feng , A. Loukas , J.M. Hawdon & P.J. Hotez (2003) Molecular characterisation of the Ancylostoma-secreted protein family from the adult stage of Ancylostoma caninum. International Journal for Parasitology 33, 897907.

Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

Journal of Helminthology
  • ISSN: 0022-149X
  • EISSN: 1475-2697
  • URL: /core/journals/journal-of-helminthology
Please enter your name
Please enter a valid email address
Who would you like to send this to? *
×

Metrics

Altmetric attention score

Full text views

Total number of HTML views: 11
Total number of PDF views: 31 *
Loading metrics...

Abstract views

Total abstract views: 199 *
Loading metrics...

* Views captured on Cambridge Core between September 2016 - 23rd September 2017. This data will be updated every 24 hours.