Plasmodium knowlesiProvides a Rapid In Vitro and In Vivo Transfection System That Enables Double-Crossover Gene Knockout Studies
| Autor: | Alan W. Thomas, Jason M. Mwenda, Hastings Ozwara, Clemens H. M. Kocken, Annemarie van der Wel, Annette L. Beetsma |
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| Rok vydání: | 2002 |
| Předmět: |
Male
Time Factors Genes Protozoan Immunology Protozoan Proteins Transfection Microbiology Apicomplexa In vivo parasitic diseases Animals Plasmodium knowlesi Crossing Over Genetic Gene knockout Genetics biology fungi biology.organism_classification Adaptation Physiological Macaca mulatta Phenotype In vitro Malaria Cell biology Circumsporozoite protein Infectious Diseases Oocytes Female Parasitology Fungal and Parasitic Infections |
| Zdroj: | Infection and Immunity. 70:655-660 |
| ISSN: | 1098-5522 0019-9567 |
| Popis: | Transfection technology for malaria parasites provides a valuable tool for analyzing gene function and correlating genotype with phenotype. Transfection models are even more valuable when appropriate animal models are available in addition to complete in vitro systems to be able to fully analyze parasite-host interactions. Here we describe the development of such a model by using the nonhuman primate malariaPlasmodium knowlesi. Blood-stage parasites were adapted to long-term in vitro culture. In vitro-adapted parasites could readapt to in vivo growth and regain wild-type characteristics after a single passage through an intact rhesus monkey.P. knowlesiparasites, either in vitro adapted or in vivo derived, were successfully transfected to generate circumsporozoite protein (CSP) knockout parasites by double-crossover mechanisms. In vitro-transfected and cloned CSP knockout parasites were derived in a time span of only 18 days. Microscopic evaluation of developing oocysts from mosquitoes that had fed on CSP knockout parasites confirmed the impairment of sporozoite formation observed inP. bergheiCSP knockout parasites. TheP. knowlesimodel currently is the only malaria system that combines rapid and precise double-crossover genetic manipulation procedures with complete in vitro as well as in vivo possibilities. This allows for full analysis ofP. knowlesigenotype-phenotype relationships and host-parasite interactions in a system closely related to humans. |
| Databáze: | OpenAIRE |
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