Driving midgut‐specific expression and secretion of a foreign protein in transgenic mosquitoes with AgAper1 regulatory elements

The Anopheles gambiae adult peritrophic matrix protein 1 (AgAper1) regulatory elements were used to drive the expression of phospholipase A2 (PLA2), a protein known to disrupt malaria parasite development in mosquitoes. These AgAper1 regulatory elements were sufficient to promote the accumulation of PLA2 in midgut epithelial cells before a blood meal and its release into the lumen upon blood ingestion. Plasmodium berghei oocyst formation was reduced by ∼80% (74–91% range) in transgenic mosquitoes. Blood‐seeking behaviour and survival of AgAper1‐PLA2 transgenic mosquitoes were comparable to sibling wild‐type mosquitoes, while fertility was substantially lower. Ultrastructural studies suggest that decreased fitness is a consequence of internal damage to midgut epithelial cells.

[1]  D. Barnard Biology of Disease Vectors , 2005 .

[2]  M. Jacobs-Lorena,et al.  Storage and secretion of the peritrophic matrix protein Ag‐Aper1 and trypsin in the midgut of Anopheles gambiae , 2004, Insect molecular biology.

[3]  L. Alphey,et al.  Editorial: Genetic control of vector populations: an imminent prospect , 2004, Tropical medicine & international health : TM & IH.

[4]  J. Wang,et al.  Fitness of Anopheline Mosquitoes Expressing Transgenes That Inhibit Plasmodium Development , 2004, Genetics.

[5]  Y. Tsujimoto,et al.  PLA2 activity is required for nuclear shrinkage in caspase-independent cell death , 2003, The Journal of cell biology.

[6]  Michael A. Riehle,et al.  Towards genetic manipulation of wild mosquito populations to combat malaria: advances and challenges , 2003, Journal of Experimental Biology.

[7]  A. Ghosh,et al.  The peritrophic matrix limits the rate of digestion in adult Anopheles stephensi and Aedes aegypti mosquitoes. , 2003, Journal of insect physiology.

[8]  A. James,et al.  Engineering Plasmodium-refractory phenotypes in mosquitoes. , 2003, Trends in parasitology.

[9]  A. Ghosh,et al.  Bee Venom Phospholipase Inhibits Malaria Parasite Development in Transgenic Mosquitoes* , 2002, The Journal of Biological Chemistry.

[10]  Chikashi Nakamura,et al.  Design and Activity of Antimicrobial Peptides against Sporogonic-Stage Parasites Causing Murine Malarias , 2002, Antimicrobial Agents and Chemotherapy.

[11]  A. Ghosh,et al.  Transgenic anopheline mosquitoes impaired in transmission of a malaria parasite , 2002, Nature.

[12]  F. Catteruccia,et al.  piggyBac-mediated Germline Transformation of the Malaria Mosquito Anopheles stephensi Using the Red Fluorescent Protein dsRED as a Selectable Marker* , 2002, The Journal of Biological Chemistry.

[13]  Xiao-Fan Wang,et al.  Transgenic anopheline mosquitoes impaired in transmission of a malaria parasite , 2002 .

[14]  J. Eaton,et al.  Delayed oxidant‐induced cell death involves activation of phospholipase A2 , 2001, FEBS letters.

[15]  T. K. Stevens,et al.  Germline transformation of the malaria vector, Anopheles gambiae, with the piggyBac transposable element , 2001, Insect molecular biology.

[16]  J. Ribeiro,et al.  A snake venom phospholipase A(2) blocks malaria parasite development in the mosquito midgut by inhibiting ookinete association with the midgut surface. , 2001, The Journal of experimental biology.

[17]  A. Raikhel,et al.  Efficient transformation of the yellow fever mosquito Aedes aegypti using the piggyBac transposable element vector pBac[3xP3-EGFP afm]. , 2001, Insect biochemistry and molecular biology.

[18]  M. Jacobs-Lorena,et al.  Targeting Plasmodium ligands on mosquito salivary glands and midgut with a phage display peptide library , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[19]  A. James,et al.  Robust gut-specific gene expression in transgenic Aedes aegypti mosquitoes. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[20]  Andrea Crisanti,et al.  Stable germline transformation of the malaria mosquito Anopheles stephensi , 2000, Nature.

[21]  A. James,et al.  Virus-expressed, recombinant single-chain antibody blocks sporozoite infection of salivary glands in Plasmodium gallinaceum-infected Aedes aegypti. , 2000, The American journal of tropical medicine and hygiene.

[22]  M. Delepierre,et al.  From noxiustoxin to Shiva-3, a peptide toxic to the sporogonic development of Plasmodium berghei. , 1998, Toxicon : official journal of the International Society on Toxinology.

[23]  Zhicheng Shen,et al.  A type I peritrophic matrix protein from the malaria vector Anopheles gambiae binds to chitin. Cloning, expression, and characterization. , 1998, The Journal of biological chemistry.

[24]  A. James,et al.  Stable transformation of the yellow fever mosquito, Aedes aegypti, with the Hermes element from the housefly. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[25]  A. James,et al.  Mariner transposition and transformation of the yellow fever mosquito, Aedes aegypti. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[26]  M. Jacobs-Lorena,et al.  Trypsin and aminopeptidase gene expression is affected by age and food composition in Anopheles gambiae. , 1996, Insect biochemistry and molecular biology.

[27]  P. Billingsley,et al.  The role of the mosquito peritrophic membrane in bloodmeal digestion and infectivity of Plasmodium species. , 1992, The Journal of parasitology.

[28]  H. Verheij,et al.  Cloning, expression, and purification of porcine pancreatic phospholipase A2 and mutants. , 1991, Methods in enzymology.

[29]  R. Gass Influences of blood digestion on the development of Plasmodium gallinaceum (Brumpt) in the midgut of Aedes aegypti (L.). , 1977, Acta tropica.