Stability and loss of a virus resistance phenotype over time in transgenic mosquitoes harbouring an antiviral effector gene

Transgenic Aedes aegypti were engineered to express a virus-derived, inverted repeat (IR) RNA in the mosquito midgut to trigger RNA interference (RNAi) and generate resistance to dengue virus type 2 (DENV2) in the vector. Here we characterize genotypic and phenotypic stabilities of one line, Carb77,...

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Bibliographic Details
Published in:Insect molecular biology 2009-10, Vol.18 (5), p.661-672
Main Authors: Franz, A.W.E, Sanchez-Vargas, I, Piper, J, Smith, M.R, Khoo, C.C.H, James, A.A, Olson, K.E
Format: Article
Language:English
Subjects:
Aedes aegypti
Animals
Animals, Genetically Modified
Antiviral Agents - metabolism
Base Sequence
Culicidae
Culicidae - genetics
Culicidae - virology
Dengue virus
Dengue Virus - physiology
Dengue virus type 2
Female
Genes, Insect
Genotype
Inheritance Patterns - genetics
Molecular Sequence Data
mosquito
Mutagenesis, Insertional - genetics
Phenotype
resistance phenotype
RNA interference
Time Factors
transgene integration site
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Summary:Transgenic Aedes aegypti were engineered to express a virus-derived, inverted repeat (IR) RNA in the mosquito midgut to trigger RNA interference (RNAi) and generate resistance to dengue virus type 2 (DENV2) in the vector. Here we characterize genotypic and phenotypic stabilities of one line, Carb77, between generations G₉ and G₁₇. The anti-DENV2 transgene was integrated at a single site within a noncoding region of the mosquito genome. The virus resistance phenotype was strong until G₁₃ and suppressed replication of different DENV2 genotypes. From G₁₄-G₁₇ the resistance phenotype to DENV2 became weaker and eventually was lost. Although the sequence of the transgene was not mutated, expression of the IR effector RNA was not detected and the Carb77 G₁₇ mosquitoes lost their ability to silence the DENV2 genome.
ISSN:0962-1075
1365-2583
DOI:10.1111/j.1365-2583.2009.00908.x