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Successful Genetic Transfection of the Colonic Protistan Parasite Blastocystis for Reliable Expression of Ectopic Genes

Li, Feng-Jun, Tsaousis, Anastasios D, Purton, Tracey J, Chow, Vincent T. K., He, Cynthia Y., Tan, Kevin S. W. (2019) Successful Genetic Transfection of the Colonic Protistan Parasite Blastocystis for Reliable Expression of Ectopic Genes. Scientific Reports, 9 . E-ISSN 2045-2322. (doi:10.1038/s41598-019-39094-5)

Abstract

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.

Item Type: Article
DOI/Identification number: 10.1038/s41598-019-39094-5
Subjects: Q Science > QR Microbiology
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: Anastasios Tsaousis
Date Deposited: 28 Feb 2019 12:21 UTC
Last Modified: 03 Jun 2019 09:30 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/72799 (The current URI for this page, for reference purposes)
Tsaousis, Anastasios D: https://orcid.org/0000-0002-5424-1905
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