Successful Genetic Transfection of the Colonic Protistan Parasite Blastocystis for Reliable Expression of Ectopic Genes
| Autor: | Cynthia Y. He, Tracey J Purton, Kevin S. W. Tan, Feng-Jun Li, Vincent T. K. Chow, Anastasios D. Tsaousis |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Untranslated region Polyadenylation Colon lcsh:Medicine Blastocystis Infections Gene delivery Legumain Article Feces 03 medical and health sciences 0302 clinical medicine Animals Humans Parasite hosting lcsh:Science Promoter Regions Genetic Gene Genetics Blastocystis Multidisciplinary biology lcsh:R Gene Transfer Techniques biology.organism_classification Gastrointestinal Microbiome 3. Good health QR Cysteine Endopeptidases 030104 developmental biology Gene Expression Regulation Host-Pathogen Interactions biology.protein lcsh:Q Ectopic expression 5' Untranslated Regions 030217 neurology & neurosurgery |
| Zdroj: | Scientific Reports Scientific Reports, Vol 9, Iss 1, Pp 1-11 (2019) |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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