Hostname: page-component-7dd5485656-2pp2p Total loading time: 0 Render date: 2025-10-30T23:18:49.687Z Has data issue: false hasContentIssue false
  • English
  • Français

RNA interference in schistosomes: machinery and methodology

Published online by Cambridge University Press:  21 September 2009

GREICE KRAUTZ-PETERSON ,
RITA BHARDWAJ ,
ZAHRA FAGHIRI ,
CIBELE A. TARARAM  and
PATRICK J. SKELLY
GREICE KRAUTZ-PETERSON
Affiliation:
Molecular Helminthology Laboratory, Division of Infectious Diseases, Department of Biomedical Sciences, Tufts University, Cummings School of Veterinary Medicine, Grafton, MA01536, USA
RITA BHARDWAJ
Affiliation:
Molecular Helminthology Laboratory, Division of Infectious Diseases, Department of Biomedical Sciences, Tufts University, Cummings School of Veterinary Medicine, Grafton, MA01536, USA
ZAHRA FAGHIRI
Affiliation:
Molecular Helminthology Laboratory, Division of Infectious Diseases, Department of Biomedical Sciences, Tufts University, Cummings School of Veterinary Medicine, Grafton, MA01536, USA
CIBELE A. TARARAM
Affiliation:
Centro de Biotecnologia, Instituto Butantan, Av. Vital Brasil, 1500, 05503-900São Paulo, SP, Brazil
PATRICK J. SKELLY*
Affiliation:
Molecular Helminthology Laboratory, Division of Infectious Diseases, Department of Biomedical Sciences, Tufts University, Cummings School of Veterinary Medicine, Grafton, MA01536, USA
*
*Corresponding author: Molecular Helminthology Laboratory, Division of Infectious Diseases, Department of Biomedical Sciences, Tufts University, Cummings School of Veterinary Medicine, Grafton, MA01536, USA. Tel: 508-887-4348. Fax: 508-839-7911. E-mail: Patrick.Skelly@Tufts.edu
Get access Rights & Permissions [Opens in a new window]

Summary

RNA interference (RNAi) is a potent gene silencing process that is playing an increasingly important role in investigations of gene function in schistosomes. Here we review what is known about the process in these parasites and provide an update on the methodology and machinery of RNAi. Data are presented to demonstrate that: (1) not all schistosome genes can be suppressed to the same extent, using the methods employed here; (2) while there is variation in the level of suppression achieved for one target gene (SmAP) in adult parasites, all individuals exhibit robust (>80%) suppression; (3) short interfering RNAs (siRNAs) can effect suppression when delivered by soaking (and not just via electroporation, as reported previously); (4) Male/female adult pairs need not be separated prior to siRNA delivery by electroporation for effective gene suppression in both genders and (5) electroporation of siRNAs in medium is as efficient as in commercial electroporation buffer. Regarding the machinery of RNAi in schistosomes, a homologue of the C. elegans multi-membrane spanning, RNA importing protein SID-1 is identified in silico. The gene encoding this protein contains 21 exons and spans over 50 kb to potentially encode a 115,556 Mr protein (SmSID-1). These analyses, and a review of the literature, permit us to derive and present here a draft of potential RNAi pathways in schistosomes.

Keywords

Gene suppressionDicerRISCsiRNA

Information

Type
Research Article
Information
Parasitology , Volume 137 , Special Issue 3: Development and applications of cestode and trematode laboratory models , March 2010 , pp. 485 - 495
Copyright
Copyright © Cambridge University Press 2009

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Article purchase

Temporarily unavailable

References

REFERENCES

Aagaard, L. and Rossi, J. J. (2007). RNAi therapeutics: principles, prospects and challenges. Advances in Drug Delivery Reviews 59, 75–86.CrossRefGoogle ScholarPubMed
Aigner, A. (2006). Gene silencing through RNA interference (RNAi) in vivo: strategies based on the direct application of siRNAs. Journal of Biotechnology 124, 12–25.CrossRefGoogle ScholarPubMed
Berriman, M., Haas, B. J., Loverde, P. T., Wilson, R. A., Dillon, G. P., Cerqueira, G. C., Mashiyama, S. T., Al-Lazikani, B., Andrade, L. F., Ashton, P. D., Aslett, M. A., Bartholomeu, D. C., Blandin, G., Caffrey, C. R., Coghlan, A., Coulson, R., Day, T. A., Delcher, A., Demarco, R., Djikeng, A., Eyre, T., Gamble, J. A., Ghedin, E., Gu, Y., Hertz-Fowler, C., Hirai, H., Hirai, Y., Houston, R., Ivens, A., Johnston, D. A., Lacerda, D., Macedo, C. D., Mcveigh, P., Ning, Z., Oliveira, G., Overington, J. P., Parkhill, J., Pertea, M., Pierce, R. J., Protasio, A. V., Quail, M. A., Rajandream, M. A., Rogers, J., Sajid, M., Salzberg, S. L., Stanke, M., Tivey, A. R., White, O., Williams, D. L., Wortman, J., Wu, W., Zamanian, M., Zerlotini, A., Fraser-Liggett, C. M., Barrell, B. G. and El-Sayed, N. M. (2009). The genome of the blood fluke Schistosoma mansoni. Nature 460, 352–358.CrossRefGoogle ScholarPubMed
Chua, J. H., Armugam, A. and Jeyaseelan, K. (2009). MicroRNAs: Biogenesis, function and applications. Current Opinion in Molecular Therapeutics 11, 189–199.Google ScholarPubMed
Conte, D. Jr. and Mello, C. C. (2003). RNA interference in Caenorhabditis elegans. Current Protocols in Molecular Biology Chapter 26, Unit 26 23.CrossRefGoogle ScholarPubMed
Correnti, J. M., Brindley, P. J. and Pearce, E. J. (2005). Long-term suppression of cathepsin B levels by RNA interference retards schistosome growth. Molecular and Biochemical Parasitology 143, 209–215.CrossRefGoogle ScholarPubMed
Delcroix, M., Sajid, M., Caffrey, C. R., Lim, K. C., Dvorak, J., Hsieh, I., Bahgat, M., Dissous, C. and Mckerrow, J. H. (2006). A multienzyme network functions in intestinal protein digestion by a platyhelminth parasite. Journal of Biological Chemistry 281, 39316–39329.CrossRefGoogle ScholarPubMed
Dinguirard, N. and Yoshino, T. P. (2006). Potential role of a CD36-like class B scavenger receptor in the binding of modified low-density lipoprotein (acLDL) to the tegumental surface of Schistosoma mansoni sporocysts. Molecular and Biochemical Parasitology 146, 219–230.CrossRefGoogle Scholar
Dusanic, D. G. (1959). Histochemical observations of alkaline phosphatase in Schistosoma mansoni. Journal of Infectious Diseases 105, 1–8.CrossRefGoogle ScholarPubMed
Duxbury, M. S., Ashley, S. W. and Whang, E. E. (2005). RNA interference: a mammalian SID-1 homologue enhances siRNA uptake and gene silencing efficacy in human cells. Biochemical and Biophysical Research Communications 331, 459–463.CrossRefGoogle ScholarPubMed
Faghiri, Z. and Skelly, P. J. (2009). The role of tegumental aquaporin from the human parasitic worm, Schistosoma mansoni, in osmoregulation and drug uptake. FASEB Journal 23, 2780–2789.CrossRefGoogle ScholarPubMed
Feinberg, E. H. and Hunter, C. P. (2003). Transport of dsRNA into cells by the transmembrane protein SID-1. Science 301, 1545–1547.CrossRefGoogle ScholarPubMed
Freitas, T. C., Jung, E. and Pearce, E. J. (2007). TGF-beta signaling controls embryo development in the parasitic flatworm Schistosoma mansoni. PLoS Pathogens 3, e52.CrossRefGoogle ScholarPubMed
Gomes, M. S., Cabral, F. J., Jannotti-Passos, L. K., Carvalho, O., Rodrigues, V., Baba, E. H. and Sa, R. G. (2009). Preliminary analysis of miRNA pathway in Schistosoma mansoni. Parasitology International 58, 61–68.CrossRefGoogle ScholarPubMed
Halton, D. W. (1967). Studies on phosphatase activity in trematoda. Journal of Parasitology 53, 46–54.CrossRefGoogle ScholarPubMed
Hoffmann, K. F. and Dunne, D. W. (2003). Characterization of the Schistosoma transcriptome opens up the world of helminth genomics. Genome Biology 5, 203.CrossRefGoogle ScholarPubMed
Janowski, B. A., Younger, S. T., Hardy, D. B., Ram, R., Huffman, K. E. and Corey, D. R. (2007). Activating gene expression in mammalian cells with promoter-targeted duplex RNAs. Nature Chemical Biology 3, 166–173.CrossRefGoogle ScholarPubMed
Krautz-Peterson, G., Camargo, S., Huggel, K., Verrey, F., Shoemaker, C. B. and Skelly, P. J. (2007 a). Amino acid transport in schistosomes: characterization of the permease heavy chain SPRM1hc. Journal of Biological Chemistry 282, 21767–21775.CrossRefGoogle ScholarPubMed
Krautz-Peterson, G., Radwanska, M., Ndegwa, D., Shoemaker, C. B. and Skelly, P. J. (2007 b). Optimizing gene suppression in schistosomes using RNA interference. Molecular and Biochemical Parasitology 153, 194–202.CrossRefGoogle ScholarPubMed
Krautz-Peterson, G. and Skelly, P. J. (2008 a). Schistosoma mansoni: the dicer gene and its expression. Experimental Parasitology 118, 122–128.CrossRefGoogle ScholarPubMed
Krautz-Peterson, G. and Skelly, P. J. (2008 b). Schistosome asparaginyl endopeptidase (legumain) is not essential for cathepsin B1 activation in vivo. Molecular and Biochemical Parasitology 159, 54–58.CrossRefGoogle Scholar
Lendner, M., Doligalska, M., Lucius, R. and Hartmann, S. (2008). Attempts to establish RNA interference in the parasitic nematode Heligmosomoides polygyrus. Molecular and Biochemical Parasitology 161, 21–31.CrossRefGoogle ScholarPubMed
Liu, F., Zhou, Y., Wang, Z. Q., Lu, G., Zheng, H., Brindley, P. J., Mcmanus, D. P., Blair, D., Zhang, Q. H., Zhong, Y., Wang, S., Han, Z. G. and Chen, Z. (2009). The Schistosoma japonicum genome reveals features of host-parasite interplay. Nature 460, 345–351.Google Scholar
Liu, J., Carmell, M. A., Rivas, F. V., Marsden, C. G., Thomson, J. M., Song, J. J., Hammond, S. M., Joshua-Tor, L. and Hannon, G. J. (2004). Argonaute2 is the catalytic engine of mammalian RNAi. Science 305, 1437–1441.CrossRefGoogle ScholarPubMed
Liu, X., Fortin, K. and Mourelatos, Z. (2008). MicroRNAs: biogenesis and molecular functions. Brain Pathology 18, 113–121.CrossRefGoogle ScholarPubMed
Livak, K. J. and Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25, 402–408.CrossRefGoogle ScholarPubMed
Morales, M. E., Rinaldi, G., Gobert, G. N., Kines, K. J., Tort, J. F. and Brindley, P. J. (2008). RNA interference of Schistosoma mansoni cathepsin D, the apical enzyme of the hemoglobin proteolysis cascade. Molecular and Biochemical Parasitology 157, 160–168.CrossRefGoogle ScholarPubMed
Morris, G. P. (1968). Fine structure of the gut epithelium of Schistosoma mansoni. Experientia 24, 480–482.CrossRefGoogle ScholarPubMed
Morris, K. V., Santoso, S., Turner, A. M., Pastori, C. and Hawkins, P. G. (2008). Bidirectional transcription directs both transcriptional gene activation and suppression in human cells. PLoS Genetetics 4, e1000258.CrossRefGoogle ScholarPubMed
Ndegwa, D., Krautz-Peterson, G. and Skelly, P. J. (2007). Protocols for gene silencing in schistosomes. Experimental Parasitology 117, 284–291.CrossRefGoogle ScholarPubMed
Okamura, K., Ishizuka, A., Siomi, H. and Siomi, M. C. (2004). Distinct roles for Argonaute proteins in small RNA-directed RNA cleavage pathways. Genes and Development 18, 1655–1666.CrossRefGoogle ScholarPubMed
Oliveira, G. (2007). The Schistosoma mansoni transcriptome: an update. Experimental Parasitology 117, 229–235.CrossRefGoogle ScholarPubMed
Pereira, T. C., Pascoal, V. D., Marchesini, R. B., Maia, I. G., Magalhaes, L. A., Zanotti-Magalhaes, E. M. and Lopes-Cendes, I. (2008). Schistosoma mansoni: evaluation of an RNAi-based treatment targeting HGPRTase gene. Experimental Parasitology 118, 619–623.CrossRefGoogle ScholarPubMed
Pratt, A. J. and Macrae, I. J. (2009). The RNA induced silencing complex: A versatile gene-silencing machine. Journal of Biological Chemistry 284, 17897–17901.CrossRefGoogle ScholarPubMed
Rinaldi, G., Morales, M. E., Alrefaei, Y. N., Cancela, M., Castillo, E., Dalton, J. P., Tort, J. F. and Brindley, P. J. (2009). RNA interference targeting leucine aminopeptidase blocks hatching of Schistosoma mansoni eggs. Molecular and Biochemical Parasitology 167, 118–126.CrossRefGoogle ScholarPubMed
Rivas, F. V., Tolia, N. H., Song, J. J., Aragon, J. P., Liu, J., Hannon, G. J. and Joshua-Tor, L. (2005). Purified Argonaute2 and an siRNA form recombinant human RISC. Nature Structural and Molecular Biology 12, 340–349.CrossRefGoogle Scholar
Saleh, M. C., Van Rij, R. P., Hekele, A., Gillis, A., Foley, E., O'farrell, P. H. and Andino, R. (2006). The endocytic pathway mediates cell entry of dsRNA to induce RNAi silencing. Nature Cell Biology 8, 793–802.CrossRefGoogle ScholarPubMed
Shih, J. D., Fitzgerald, M. C., Sutherlin, M. and Hunter, C. P. (2009). The SID-1 double-stranded RNA transporter is not selective for dsRNA length. RNA 15, 384–390.CrossRefGoogle Scholar
Silk, M. H., Spence, I. M. and Gear, J. H. (1969). Ultrastructural studies of the blood fluke – Schistosoma mansoni. I. The integument. South African Journal of Medical Science 34, 1–10.Google ScholarPubMed
Siomi, H. and Siomi, M. C. (2009). On the road to reading the RNA-interference code. Nature 457, 396–404.CrossRefGoogle ScholarPubMed
Skelly, P. J. (2006). Gene silencing in flatworms using RNA interference. In Parasitic Flatworms Molecular Biology, Biochemistry, Immunology and Physiology (ed. Maule, A. G. & Marks, N. J.), pp. 423–434. CAB International, Oxford.CrossRefGoogle Scholar
Skelly, P. J., Da'dara, A. and Harn, D. A. (2003). Suppression of cathepsin B expression in Schistosoma mansoni by RNA interference. International Journal for Parasitology 33, 363–369.CrossRefGoogle ScholarPubMed
Spence, I. M. and Silk, M. H. (1970). Ultrastructural studies of the blood fluke – Schistosoma mansoni. IV. The digestive system. South African Journal of Medical Science 35, 93–112.Google ScholarPubMed
Spence, I. M. and Silk, M. H. (1971). Ultrastructural studies of the blood fluke – Schistosoma mansoni. V. The female reproductive system – a preliminary report. South African Journal of Medical Science 36, 41–50.Google ScholarPubMed
Van Hellemond, J. J., Van Balkom, B. W. and Tielens, A. G. (2007). Schistosome biology and proteomics: progress and challenges. Experimental Parasitology 117, 267–274.CrossRefGoogle ScholarPubMed
Verjovski-Almeida, S., Demarco, R., Martins, E. A., Guimaraes, P. E., Ojopi, E. P., Paquola, A. C., Piazza, J. P., Nishiyama, M. Y. Jr., Kitajima, J. P., Adamson, R. E., Ashton, P. D., Bonaldo, M. F., Coulson, P. S., Dillon, G. P., Farias, L. P., Gregorio, S. P., Ho, P. L., Leite, R. A., Malaquias, L. C., Marques, R. C., Miyasato, P. A., Nascimento, A. L., Ohlweiler, F. P., Reis, E. M., Ribeiro, M. A., Sa, R. G., Stukart, G. C., Soares, M. B., Gargioni, C., Kawano, T., Rodrigues, V., Madeira, A. M., Wilson, R. A., Menck, C. F., Setubal, J. C., Leite, L. C. and Dias-Neto, E. (2003). Transcriptome analysis of the acoelomate human parasite Schistosoma mansoni. Nature Genetics 35, 148–157.CrossRefGoogle ScholarPubMed
Whangbo, J. S. and Hunter, C. P. (2008). Environmental RNA interference. Trends in Genetics 24, 297–305.CrossRefGoogle ScholarPubMed
Wilson, R. A., Ashton, P. D., Braschi, S., Dillon, G. P., Berriman, M. and Ivens, A. (2007). ‘Oming in on schistosomes: prospects and limitations for post-genomics. Trends in Parasitology 23, 14–20.CrossRefGoogle Scholar
Winston, W. M., Molodowitch, C. and Hunter, C. P. (2002). Systemic RNAi in C. elegans requires the putative transmembrane protein SID-1. Science 295, 2456–2459.CrossRefGoogle Scholar
Xue, X., Sun, J., Zhang, Q., Wang, Z., Huang, Y. and Pan, W. (2008). Identification and characterization of novel microRNAs from Schistosoma japonicum. PLoS ONE 3, e4034.CrossRefGoogle ScholarPubMed