| 研究実績の概要 |
On this fiscal year, we tried to analyze the gene in-vivo by injecting l(2)efl-1, -2, -3, and -4 dsRNAs to mosquito thoraxes and then followed by DENV-2 injection 48 hours after dsRNA-injection. We used the same dsRNA that we transfected in CCL-125. We did preliminary experiment to see how long the dsRNA would give an effect in-vivo. We found that 72 hours after dsRNA injection, the level of l(2)efl-1 to l(2)efl-5 was suppressed. Because we needed to consider that after 72 hours dsRNA might be losing its effect, we decided to inject DENV-2 48 hours after dsRNA injection. Following dsRNA-injection, mosquitoes looked healthy with only little number of mosquitoes died. Forty eight hours afterwards, we injected mosquitoes with DENV-2 and sacrificed the mosquitoes 24, 48, and 72 hours later. Unfortunately, using qRT-PCR, we found that there was no significant difference between dsRNA-injected and control mosquitoes.
The reason that DENV-2 kept on replicating might because dsRNA was losing its effect at the time of DENV-2 injection. In the future, we will try to co-inject DENV-2 and dsRNA at the same time or to create a transgenic mosquito by injecting dsRNA at the embryo stage. We have submitted the results of in-vitro experiment to Japanese Journal of Infectious Disease for consideration.
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