RNA interference effects triggered by dsRNA and siRNA in Aedes albopictus
- Author:
WU Qixing
;
JIANG Yuting
;
LIU Lu
- Publication Type:Journal Article
- Keywords:
Aedes albopictus;
RNA interference;
siRNA;
dsRNA
- From:
China Tropical Medicine
2024;24(11):1411-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the interference effects of siRNA and dsRNA in Aedes albopictus and their impact on mosquito survival, providing reference and technical support for the study of mosquito gene function, the research and development of new insecticides, the control of vector mosquitoes, and the interruption of arboviruses transmission. Methods For genes of interest in Ae. albopictus, siRNAs targeting the desired genes were designed and commercially synthesized, while dsRNAs were obtained by amplifying the target gene fragments, followed by in vitro transcription and purification, with verification by agarose gel electrophoresis. The interfering RNA was injected into Ae. albopictus by thoracic microinjection. Then the expression of targeted genes on the 3rd day post-microinjection was detected by quantitative PCR to verify the interfering effect, and the survival curve of the mosquitoes was recorded at the same time. Results In the siRNA interference experiments targeting 10 genes with 22 siRNA pairs, only two siRNA pairs, SR-C-2 siRNA and ENR-1 siRNA, successfully interfered with the target gene expression compared to the control group, with interference rates of 58% and 33%, respectively, while the remaining siRNAs did not show interference effects, with some even causing increased expression of the target gene. In the dsRNA interference experiments, Mesh dsRNA significantly reduced the expression of mesh gene at the concentration of 3×1011 RNA molecules per mosquito, with an interference effect lasting 5 days and an interference rate of 55%-60% on the third day and 34% on the fifth-day post-injection. Mosquito survival within 3 days post-dsRNA injection was significantly higher than in the siRNA injection group. Conclusions Ae. albopictus on the fifth-day post-emergence is the most suitable to receive thoracic microinjection. dsRNA is more stable than siRNA for interference, with higher post-injection survival rates, making it more suitable for in-depth studies of mosquito gene function. Although siRNA-based gene interference is simpler and more feasible, attention should be paid to the purification method of siRNA and its homology with other genes in the mosquito species to avoid non-specific experimental results due to the off-target combinations.
- Full text:202511121027077937519.RNA interference effects triggered by dsRNA and siRNA in Aedes albopictus.pdf
