2002 Fiscal Year Final Research Report Summary
Generation of transgenic anopheline mosquitoes refractory to malaria parasite by using gene manipulation
| Project/Area Number |
13670253
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
寄生虫学(含医用動物学)
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| Research Institution | Jichi Medical School |
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Principal Investigator |
YOSHIDA Shigeto Jichi Medical School Assistant Professor, 医学部, 講師 (10296121)
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| Co-Investigator(Kenkyū-buntansha) |
HIRAI Makoto Jichi Medical School Research Associate, 医学部, 助手 (50326849)
MATSUOKA Hiroyuki Jichi Medical School Associate Professor, 医学部, 助教授 (10173816)
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| Project Period (FY) |
2001 – 2002
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| Keywords | Malaria / Anopheles stephensi / Genetic manipulation / Transgcnic / single-chain antibody / Transmission-blocking |
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| Research Abstract |
In collaboration with Prof.Crisanti, the head investigator cloned the promoter region of the Anopheles gambiae carboxypeptidase (AgCP) gene. Two plasmid constructs (cytoplasmic and secreted forms) encoding the 13.1 single-chain antibody linked to Shiva (13.1 scFv-Shiva) under the control of the AgCP promoter were made. The plasmid DNA harboring the cytoplasmic gene was injected into Anopheles stephensi embryos. Using fluorescent positive selection, 5 lines of transgenic mosquitoes were obtained. The 13.1 scFv-Shiva gene was abundantly expressed in the guts of transgenic mosquitoes. Expression of the 13.1 scFv-Shiva was gut-specific and reached peak levels at about 3 h post-blood ingestion. The AgCP promoter can be used to drive the expression of genes that hinder parasite development in the mosquito gut. To measure the consequences of the 13.1 scFv-Shiva transgene expression on the parasite development, we fed control and transgenic mosquitoes on the same infected mouse and measured the numbers of oocysts formed Oocyst formation in transgenic mosquitoes was indistinguishable from that in wild type mosquitoes. Another line expressing the secreted form of 13.1 scFv-Shiva is in progress of examining.
Prof.Crisanti's group showed that bee venom phospholipsase inhibits malaria parasites development in transgenic mosquitoes (JBC 2002). They also showed that genetic modification of mosquitoes offers exciting possibility for controlling malaria, but success will depend on how transmission affects the fitness of modified insects (Science 2003).
Prof.Sinden's group joined the projects on genomic sequence of rodent malaria parasites. The sequence information provides insight into Plasmodium biology and disease (Nature 2002).
Thus, we have established technology and facility for generating transgenic mosquitoes. This work represents a significant step toward the development of molecular genetic approaches to the control of vector competence in malaria transmission.
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[Publications] Tomas AM, Margos G, Dimopoulos G, van Lin LH, de Koning-Ward TF, Sinha R, Lupetti P, Beetsma AL, Rodriguez MC, Karras M, Hager A, Mendoza J, Butcher GA, Kafatos F, Janse CJ, Waters AP, Sinden RE.: "P25 and P28 proteins of the malaria ookinete surface have multiple and partially redundant functions"EMBO J.. 20. 3975-3983 (2001)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Carlton JM, Angiuoli SV, Suh BB, Kooij TW, Pertea M, Silva JC, Ermolaeva MD, Allen JE, Selengut JD, Koo HL, Peterson JD, Pop M, Kosack DS, Shumway MF, Bidwell SL, Shallom SJ, van Aken SE, Riedmuller SB, Feldblyum TV, Cho JK, Quackenbush J, Sedegah M, Shoaibi A, Cummings LM, Florens L, Yates JR, Raine JD, Sinden RE, Harris MA, Cunningham DA, Preiser PR, Bergman LW, Vaidya AB, van Lin LH, Janse CJ, Waters AP, Smith HO, White OR, Salzberg SL, Venter JC, Fraser CM, Hoffman SL, Gardner MJ, Carucci DJ.: "Genome sequence and comparative analysis of the model rodent malaria parasite Plasmodium yoelii yoelii"Nature. 419. 512-519 (2002)
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「研究成果報告書概要(欧文)」より
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[Publications] Flrens L, Washburn MP, Raine JD, Anthony RM, Grainger M, Haynes JD, Moch JK, Muster N, Sacci JB, Tabb DL, Witney AA, Wolters D, Wu Y, Gardner MJ, Holder AA, Sinden RE, Yates JR, Carucci DJ.: "A proteomic view of the Plasmodium falciparum life cycle"Nature. 419. 520-526 (2002)
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「研究成果報告書概要(欧文)」より
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[Publications] Alphey L, Beard CB, Billingsley P, Coetzee M, Crisanti A, Curtis C, Eggleston P, Godfray C, Hemingway J, Jacobs-Lorena M, James AA, Kafatos FC, Mukwaya LG, Paton M, Powell JR, Schneider W, Scott TW, Sina B, Sinden R, Sinkins S, Spielman A, Toure Y, Collins FH.: "Malaria control with genetically manipulated insect vectors"Science. 298. 119-121 (2002)
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「研究成果報告書概要(欧文)」より
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