Developmental sensitivity in schistosoma mansoni to puromycin to establish drug selection of transgenic schistosomes

Hong Bin Yan, Michael J. Smout, Chuan Ju, Anne E. Folley, Danielle E. Skinner, Victoria H. Mann, Alex Loukas, Wei Hu, Paul J. Brindley, Gabriel Rinaldi*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

8 Citations (SciVal)


Schistosomiasis is considered the most important disease caused by helminth parasites, in terms of morbidity and mortality. Tools to facilitate gain- and loss-of-function approaches can be expected to precipitate the discovery of novel interventions, and drug selection of transgenic schistosomes would facilitate the establishment of stable lines of engineered parasites. Sensitivity of developmental stages of schistosomes to the aminonucleoside antibiotic puromycin was investigated. For the schistosomulum and sporocyst stages, viability was quantified by fluorescence microscopy following dual staining with fluorescein diacetate and propidium iodine. By 6 days in culture, the 50% lethal concentration (LC 50) for schistosomula was 19 μg/ml whereas the sporocysts were 45-fold more resilient. Puromycin potently inhibited the development of in vitro-laid eggs (LC 50, 68 ng/ml) but was less effective against liver eggs (LC 50, 387 μg/ml). Toxicity for adult stages was evaluated using the xCELLigence-based, real-time motility assay (xWORM), which revealed LC 50s after 48 h of 4.9 and 17.3 μg/ml for male and female schistosomes, respectively. Also, schistosomula transduced with pseudotyped retrovirus encoding the puromycin resistance marker were partially rescued when cultured in the presence of the antibiotic. Together, these findings will facilitate selection on puromycin of transgenic schistosomes and the enrichment of cultures of transgenic eggs and sporocysts to facilitate the establishment of schistosome transgenic lines. Streamlining schistosome transgenesis with drug selection will open new avenues to understand parasite biology and hopefully lead to new interventions for this neglected tropical disease.

Original languageEnglish
Article numbere02568-17
JournalAntimicrobial Agents and Chemotherapy
Issue number8
Publication statusPublished - Aug 2018
Externally publishedYes


  • Antibiotic susceptibility
  • Drug selection
  • Functional genomics
  • Puromycin
  • Schistosomiasis
  • Puromycin/pharmacology
  • Animals, Genetically Modified
  • Schistosomiasis/drug therapy
  • Fluoresceins/pharmacology
  • Male
  • Genomics/methods
  • Animals
  • Schistosoma mansoni/drug effects
  • Female


Dive into the research topics of 'Developmental sensitivity in schistosoma mansoni to puromycin to establish drug selection of transgenic schistosomes'. Together they form a unique fingerprint.

Cite this