Please use this identifier to cite or link to this item: https://doi.org/10.1038/s41598-019-39094-5
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dc.titleSuccessful Genetic Transfection of the Colonic Protistan Parasite Blastocystis for Reliable Expression of Ectopic Genes
dc.contributor.authorLi, FJ
dc.contributor.authorTsaousis, AD
dc.contributor.authorPurton, T
dc.contributor.authorChow, VTK
dc.contributor.authorHe, CY
dc.contributor.authorTan, KSW
dc.date.accessioned2019-06-03T04:20:43Z
dc.date.available2019-06-03T04:20:43Z
dc.date.issued2019-12-01
dc.identifier.citationLi, FJ, Tsaousis, AD, Purton, T, Chow, VTK, He, CY, Tan, KSW (2019-12-01). Successful Genetic Transfection of the Colonic Protistan Parasite Blastocystis for Reliable Expression of Ectopic Genes. Scientific Reports 9 (1) : 3159-. ScholarBank@NUS Repository. https://doi.org/10.1038/s41598-019-39094-5
dc.identifier.issn2045-2322
dc.identifier.issn2045-2322
dc.identifier.urihttps://scholarbank.nus.edu.sg/handle/10635/155020
dc.description.abstract© 2019, The Author(s). The microbial parasite Blastocystis colonizes the large intestines of numerous animal species and increasing evidence has linked Blastocystis infection to enteric diseases with signs and symptoms including abdominal pain, constipation, diarrhea, nausea, vomiting, and flatulence. It has also recently been reported to be an important member of the host intestinal microbiota. Despite significant advances in our understanding of Blastocystis cell biology and host-parasite interactions, a genetic modification tool is absent. In this study, we successfully established a robust gene delivery protocol for Blastocystis subtype 7 (ST7) and ectopic protein expression was further tested using a high sensitivity nano-luciferase (Nluc) reporter system, with promoter regions from several genes. Among them, a strong promoter encompassing a region upstream of the legumain 5′ UTR was identified. Using this promoter combined with the legumain 3′ UTR, which contains a conserved, precise polyadenylation signal, a robust transient transfection technique was established for the first time in Blastocystis. This system was validated by ectopic expression of proteins harbouring specific localization signals. The establishment of a robust, reproducible gene modification system for Blastocystis is a significant advance for Blastocystis research both in vitro and in vivo. This technique will spearhead further research to understand the parasite’s biology, its role in health and disease, along with novel ways to combat the parasite.
dc.publisherSpringer Nature
dc.sourceElements
dc.typeArticle
dc.date.updated2019-06-03T00:30:52Z
dc.contributor.departmentDEPT OF MICROBIOLOGY & IMMUNOLOGY
dc.contributor.departmentDEPT OF BIOLOGICAL SCIENCES
dc.description.doi10.1038/s41598-019-39094-5
dc.description.sourcetitleScientific Reports
dc.description.volume9
dc.description.issue1
dc.description.page3159-
dc.published.statePublished
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