Document Detail


Transgene-mediated suppression of the RNA interference pathway in Aedes aegypti interferes with gene silencing and enhances Sindbis virus and dengue virus type 2 replication.
MedLine Citation:
PMID:  23331493     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
RNA interference (RNAi) is the major innate antiviral pathway in Aedes aegypti that responds to replicating arboviruses such as dengue virus (DENV) and Sindbis virus (SINV). On the one hand, the mosquito's RNAi machinery is capable of completely eliminating DENV2 from Ae. aegypti. On the other, transient silencing of key genes of the RNAi pathway increases replication of SINV and DENV2, allowing the viruses to temporally overcome dose-dependent midgut infection and midgut escape barriers (MEB) more efficiently. Here we expressed Flock house virus B2 (FHV-B2) from the poly-ubiquitin (PUb) promoter in Ae. aegypti using the ΦC31 site-directed recombination system to investigate the impact of transgene-mediated RNAi pathway suppression on infections with SINV-TR339eGFP and DENV2-QR94, the latter of which has been shown to be confronted with a strong MEB in Ae. aegypti. FHV-B2 was constitutively expressed in midguts of sugar- and blood-fed mosquitoes of transgenic line PUbB2 P61. B2 over-expression suppressed RNA silencing of carboxypeptidase A-1 (AeCPA-1) in midgut tissue of PUbB2 P61 mosquitoes. Following oral challenge with SINV-TR339eGFP or DENV2-QR94, mean titres in midguts of PUbB2 P61 females were significantly higher at 7 days post-bloodmeal (pbm) than in those of nontransgenic control mosquitoes. At 14 days pbm, infection rates of carcasses were significantly increased in PubB2 P61 mosquitoes infected with SINV-TR339eGFP. Following infection with DENV2-QR94, midgut infection rates were significantly increased in the B2-expressing mosquitoes at 14 days pbm. However, B2 expression in PUbB2 P61 did not increase the DENV2-QR94 dissemination rate, indicating that the infection phenotype was not primarily controlled by RNAi.
Authors:
C C H Khoo; J B Doty; M S Heersink; K E Olson; A W E Franz
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  Insect molecular biology     Volume:  22     ISSN:  1365-2583     ISO Abbreviation:  Insect Mol. Biol.     Publication Date:  2013 Feb 
Date Detail:
Created Date:  2013-01-21     Completed Date:  2013-07-22     Revised Date:  2014-02-04    
Medline Journal Info:
Nlm Unique ID:  9303579     Medline TA:  Insect Mol Biol     Country:  England    
Other Details:
Languages:  eng     Pagination:  104-14     Citation Subset:  IM    
Copyright Information:
© 2013 Royal Entomological Society.
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MeSH Terms
Descriptor/Qualifier:
Aedes / genetics*,  virology*
Animals
Animals, Genetically Modified*
Dengue Virus / pathogenicity,  physiology*
Female
Gastrointestinal Tract / physiology
Gene Expression
Gene Silencing
Nodaviridae / genetics
Polyubiquitin / genetics
Promoter Regions, Genetic
RNA Interference
Sindbis Virus / pathogenicity,  physiology*
Transgenes
Virus Replication / genetics*
Grant Support
ID/Acronym/Agency:
AI073298-01/AI/NIAID NIH HHS; R01 AI073298/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
120904-94-1/Polyubiquitin
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