Inducible Knockdown of Plasmodium Gene Expression Using the glmS Ribozyme
| العنوان: | Inducible Knockdown of Plasmodium Gene Expression Using the glmS Ribozyme |
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| المؤلفون: | Ellen Knuepfer, Parichat Prommana, Philip J. Shaw, Sumalee Kamchonwongpaisan, Chayaphat Wongsombat, Anthony A. Holder, Yongyuth Yuthavong, Chairat Uthaipibull |
| المصدر: | PLoS ONE; Vol 8 PLoS ONE PLoS ONE, Vol 8, Iss 8, p e73783 (2013) |
| بيانات النشر: | PUBLIC LIBRARY SCIENCE, 2013. |
| سنة النشر: | 2013 |
| مصطلحات موضوعية: | Biochemistry, 0302 clinical medicine, Nucleic Acids, Gene expression, Molecular Cell Biology, Gene Knockdown Techniques, Ribozymes, 0303 health sciences, Gene knockdown, Glucosamine, Multidisciplinary, biology, Ribozyme, Gene targeting, 3. Good health, Phenotype, Essential gene, Medicine, Genetic Engineering, Research Article, Biotechnology, Science, 030231 tropical medicine, Plasmodium falciparum, Microbiology, Molecular Genetics, 03 medical and health sciences, Bacterial Proteins, Multienzyme Complexes, parasitic diseases, Parasite Groups, Genetics, Humans, RNA, Catalytic, Gene Regulation, RNA, Messenger, Biology, Microbial Pathogens, 030304 developmental biology, Reporter gene, Parasite Physiology, Thymidylate Synthase, Molecular biology, Reverse genetics, Reverse Genetics, Tetrahydrofolate Dehydrogenase, Gene Expression Regulation, biology.protein, RNA, Parasitology |
| الوصف: | Conventional reverse genetic approaches for study of Plasmodium malaria parasite gene function are limited, or not applicable. Hence, new inducible systems are needed. Here we describe a method to control P. falciparum gene expression in which target genes bearing a glmS ribozyme in the 3' untranslated region are efficiently knocked down in transgenic P. falciparum parasites in response to glucosamine inducer. Using reporter genes, we show that the glmS ribozyme cleaves reporter mRNA in vivo leading to reduction in mRNA expression following glucosamine treatment. Glucosamine-induced ribozyme activation led to efficient reduction of reporter protein, which could be rapidly reversed by removing the inducer. The glmS ribozyme was validated as a reverse-genetic tool by integration into the essential gene and antifolate drug target dihydrofolate reductase-thymidylate synthase (PfDHFR-TS). Glucosamine treatment of transgenic parasites led to rapid and efficient knockdown of PfDHFR-TS mRNA and protein. PfDHFR-TS knockdown led to a growth/arrest mutant phenotype and hypersensitivity to pyrimethamine. The glmS ribozyme may thus be a tool for study of essential genes in P. falciparum and other parasite species amenable to transfection. |
| اللغة: | English |
| تدمد: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0073783 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::0ad1df8814d826efc7c170bd32068a5dTest |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....0ad1df8814d826efc7c170bd32068a5d |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 19326203 |
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| DOI: | 10.1371/journal.pone.0073783 |