piggybac- and PhiC31-mediated genetic transformation of the Asian tiger mosquito, Aedes albopictus (Skuse).
BACKGROUND
The Asian tiger mosquito, Aedes albopictus (Skuse), is a vector of several arboviruses including dengue and chikungunya. This highly invasive species originating from Southeast Asia has travelled the world in the last 30 years and is now established in Europe, North and South America, Africa, the Middle East and the Caribbean. In the absence of vaccine or antiviral drugs, efficient mosquito control strategies are crucial. Conventional control methods have so far failed to control Ae. albopictus adequately.


RESULTS
Germline transformation of Aedes albopictus was achieved by micro-injection of embryos with a piggyBac-based transgene carrying a 3xP3-ECFP marker and an attP site, combined with piggyBac transposase mRNA and piggyBac helper plasmid. Five independent transgenic lines were established, corresponding to an estimated transformation efficiency of 2-3%. Three lines were re-injected with a second-phase plasmid carrying an attB site and a 3xP3-DsRed2 marker, combined with PhiC31 integrase mRNA. Successful site-specific integration was observed in all three lines with an estimated transformation efficiency of 2-6%.


CONCLUSIONS
Both piggybac- and site-specific PhiC31-mediated germline transformation of Aedes albopictus were successfully achieved. This is the first report of Ae. albopictus germline transformation and engineering, a key step towards studying and controlling this species using novel molecular techniques and genetic control strategies.
Labbé
Geneviève M C
GM
Oxitec Limited, Oxford, United Kingdom.
Nimmo
Derric D
DD
Alphey
Luke
L
D000330:Aedes / Q000235:genetics*; D000818:Animals; D030801:Animals, Genetically Modified; D001287:Attachment Sites, Microbiological; D005260:Female; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D019426:Integrases / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D008845:Microinjections / Q000379:methods; D010957:Plasmids; D011995:Recombination, Genetic; D014170:Transformation, Genetic; D019895:Transposases / Q000235:genetics; D000097573:Red Fluorescent Protein
D049452:Green Fluorescent Proteins; D019426:Integrases; D008164:Luminescent Proteins; D019895:Transposases; D000097573:Red Fluorescent Protein; C485184:enhanced green fluorescent protein
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