In vivo functional genomic studies of sterol carrier protein-2 gene in the yellow fever mosquito

• Published in: PloS one 2011-03, Vol.6 (3), p.e18030-e18030
• Main Authors: Peng, Rong, Maklokova, Vilena I, Chandrashekhar, Jayadevi H, Lan, Que
• Format: Article
• Language: English
• Subjects: Aedes; Aedes - genetics; Aedes - growth & development; Aedes aegypti; Amino acids; Animals; Apis mellifera; Aquatic insects; beta-Galactosidase - metabolism; Biology; Carrier Proteins - genetics; Carrier Proteins - metabolism; Culicidae; Deoxyribonucleic acid; Diptera; DNA; DNA - genetics; Domestication; Drosophila melanogaster; Eggs; Embryos; Endocytosis; Expression vectors; Extrachromosomal DNA; Female; Females; Fertility; Fever; Gene expression; Gene Expression Regulation; Gene Knockdown Techniques; Gene Transfer Techniques; Genes; Genes, Insect - genetics; Genetic aspects; Genetic engineering; Genetic research; Genetic vectors; Genetic Vectors - genetics; Genomics; Genomics - methods; Health aspects; Heat-Shock Response - genetics; In vivo methods and tests; Insect Proteins - genetics; Insect Proteins - metabolism; Insects; Laboratories; Larva - metabolism; Larvae; Metabolism; Microinjection; Mosquitoes; Mutation; Oocytes - cytology; Oocytes - metabolism; Proteins; Reproduction; RNA, Messenger - genetics; RNA, Messenger - metabolism; RNA, Small Interfering - metabolism; Sex ratio; siRNA; Stegomyia; Steroids (Organic compounds); Sterols; Transfection; Vector-borne diseases; Vitellogenesis; Yellow fever; Yellow Fever - genetics; Yellow Fever - parasitology
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0018030

A simple and efficient DNA delivery method to introduce extrachromosomal DNA into mosquito embryos would significantly aid functional genomic studies. The conventional method for delivery of DNA into insects is to inject the DNA directly into the embryos. Taking advantage of the unique aspects of mosquito reproductive physiology during vitellogenesis and an in vivo transfection reagent that mediates DNA uptake in cells via endocytosis, we have developed a new method to introduce DNA into mosquito embryos vertically via microinjection of DNA vectors in vitellogenic females without directly manipulating the embryos. Our method was able to introduce inducible gene expression vectors transiently into F0 mosquitoes to perform functional studies in vivo without transgenic lines. The high efficiency of expression knockdown was reproducible with more than 70% of the F0 individuals showed sufficient gene expression suppression (