Designing an Artificial Intronic Small RNA Expression Cassette
4:22
Generating a rAaeDV Construct by Cloning a miRNA or shRNA Expression Cassette into a Plasmid Backbone
5:19
Transfecting C6/36 Cells with rAaeDV Plasmids
7:33
Harvesting rAaeDV Virions from Transfected C6/36 Cells
8:53
Mosquito Transduction
10:10
Results: Overexpression and Knockdown Efficiency of rAaeDV Vectors
11:14
Conclusion
Transkript
The overall goal of this procedure is to analyze the gene function of mosquito larvae, using recombinant mosquito densovirus as a gene delivery. This method can help answer key questions in viral-based delivery systems, such as how to overcome the
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We report using an artificial intronic small RNA expression strategy to develop a non-defective recombinant Aedes aegypti densovirus (AaeDV) in vivo delivery system. A detailed procedure for the construction, packaging, and quantitative analysis of the rAaeDV vectors as well as for larval infection is described.