Transgene expression in Schistosoma mansoni: introduction of RNA into schistosomula by electroporation.

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  • Author(s): Correnti JM;Correnti JM; Pearce EJ
  • Source:
    Molecular and biochemical parasitology [Mol Biochem Parasitol] 2004 Sep; Vol. 137 (1), pp. 75-9.
  • Publication Type:
    Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: Elsevier/North-Holland Biomedical Press Country of Publication: Netherlands NLM ID: 8006324 Publication Model: Print Cited Medium: Print ISSN: 0166-6851 (Print) Linking ISSN: 01666851 NLM ISO Abbreviation: Mol Biochem Parasitol Subsets: MEDLINE
    • Publication Information:
      Original Publication: Amsterdam, Elsevier/North-Holland Biomedical Press.
    • Subject Terms:
      Electroporation* ; Gene Expression* ; Transgenes*; RNA/*metabolism ; Schistosoma mansoni/*genetics; Animals ; Genes, Reporter ; Luciferases/genetics ; Luciferases/metabolism ; RNA/genetics
    • Abstract:
      Despite their significance in human and veterinary medicine, and the ability to maintain the parasites in the mouse, relatively little functional detail is available regarding the biology of schistosomes. This deficit is due largely to the lack of well-developed molecular tools for manipulating gene expression in these parasites. Here, we describe an electroporation protocol that provides a routine approach for efficiently introducing nucleic acids into schistosomes. Using luciferase-encoding RNA for electroporation, and luciferase activity as a read-out, we established 400 microg/ml of RNA, and a 20 ms pulse at 125 V using a square wave electroporation generator to be optimal for electroporating schistosomes. Under these conditions schistosomula from 1 hr to 18 hr old could be successfully electroporated, the majority of parasites within a population expressed the introduced RNA, and acute mortality was negligible. Electroporation, as described here, makes possible experimental studies using transiently expressed constitutively active and/or dominant negative mutant proteins, etc. In addition, the finding that electroporation can be used to introduce RNA into schistosomula raises the possibility of using this approach to introduce either DNA constructs or dsRNA sequences, both of which might be expected to have longer-term, ideally inheritable, effects.
    • Grant Information:
      N01-AI-55270 United States AI NIAID NIH HHS
    • Accession Number:
      63231-63-0 (RNA)
      EC 1.13.12.- (Luciferases)
    • Publication Date:
      Date Created: 20040729 Date Completed: 20050225 Latest Revision: 20071114
    • Publication Date:
      20250114
    • Accession Number:
      10.1016/j.molbiopara.2004.04.015
    • Accession Number:
      15279953