Development of a new Sindbis virus transducing system and its characterization in three Culicine mosquitoes and two Lepidopteran species.
Alphavirus transducing systems (ATSs) are alphavirus-based tools for expressing genes in insects. Here we describe an ATS (5'dsMRE16ic) based entirely on Sindbis MRE16 virus. GFP expression was used to characterize alimentary tract infections and dissemination in three Culicine and two Lepidopteran species. Following per os infection, 5'dsMRE16ic-EGFP efficiently infected Aedes aegypti and Culex tritaeniorhynchus, but not Culex pipiens pipiens. Ae. aegypti clearly showed accumulation of green fluorescent protein (GFP) in the posterior midgut and foregut/midgut junction within 2-3 days postinfection. Following parenteral infection of larvae, Bombyx mori had extensive GFP expression in larvae and adults, but Manduca sexta larvae were mostly resistant. 5'dsMRE16ic should be a valuable tool for gene expression in several important insect species that are otherwise difficult to manipulate genetically.
Foy
B D
BD
Arthropod-Borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523, USA. brian.foy@colostate.edu
Myles
K M
KM
Pierro
D J
DJ
Sanchez-Vargas
I
I
Uhlírová
M
M
Jindra
M
M
Beaty
B J
BJ
Olson
K E
KE
D000818:Animals; D009033:Culicidae / Q000235:genetics* / Q000821:virology; D017931:DNA Primers; D004064:Digestive System / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000378:metabolism; D009036:Moths / Q000235:genetics* / Q000821:virology; D010957:Plasmids / Q000235:genetics; D012845:Sindbis Virus; D014161:Transduction, Genetic / Q000379:methods*
D017931:DNA Primers; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins