Biolistic transformation of Schistosoma mansoni: Studies with modified reporter-gene constructs containing regulatory regions of protease genes

Mol Biochem Parasitol. 2010 Mar;170(1):37-40. doi: 10.1016/j.molbiopara.2009.11.001. Epub 2009 Nov 13.

Abstract

Biolistics of the flatworm parasite Schistosoma mansoni facilitates the accurate spatial expression of transgenes under the control of gene-specific promoter elements. To improve transgene expression, either in the number of positive worms and/or an increased transgene signal per worm, we tested plasmid constructs incorporating 5' and 3' gene-specific genomic fragments, and parts of the open reading frame for two S. mansoni proteases, cathepsins F and D (SmCF and SmCD). GFP-expression was gut-localized, a novel finding for SmCD and consistent with previous data for SmCF. The mCherry fluorescent protein can also operate as a reporter. Though certain constructs imparted stronger and better distributed signals per positive worm, the low yields throughout (1-5% positive per experiment) precluded further quantifications of improvement. Electroporation of the same constructs was also weakly efficient (1-10% positives per experiment). However, reporter signals were found in tissues other than the gut, which may represent dysregulated transcription.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biolistics
  • Cathepsin D / genetics
  • Cathepsin D / metabolism
  • Cathepsin F / genetics
  • Cathepsin F / metabolism
  • Gene Expression Regulation
  • Genes, Reporter*
  • Helminth Proteins / genetics*
  • Helminth Proteins / metabolism
  • Peptide Hydrolases / genetics*
  • Peptide Hydrolases / metabolism
  • Regulatory Sequences, Nucleic Acid*
  • Schistosoma mansoni / genetics*
  • Schistosoma mansoni / metabolism
  • Transformation, Genetic*

Substances

  • Helminth Proteins
  • Peptide Hydrolases
  • Cathepsin F
  • Cathepsin D