Team:SSHS-Shenzhen/Experiments
Experimental summary
Short interfering RNAs (siRNAs) and their corresponding shRNAs were designed based on the mRNA sequences of their target genes. siRNAs were synthesized by a bio-company, shRNAs were produced by in vitro transcription. The efficiency of both siRNAs and shRNAs in mediating RNAi in Phyllotreta striolata were examined. Experimental results show that both siRNAs and shRNA could successfully silence their target genes, which was demonstrated by the survival rate decrease after siRNA or shRNA treatment. Results also show that GC percentage of the total siRNA/shRNA, the 5’ end of the siRNA/shRNA, the 3’ end of the siRNA/shRNA affect the RNAi efficiency.
In addition, we tested the attraction effect of sucrose and lemon yellow on P. S. Results show that sucrose plus lemon yellow has the best attraction effect.
Project-specific biosafety
In our project, we work with Phyllotreta striolata, which is not on the white list. We submitted check-in form about using Phyllotreta striolata in our project, and got approved by the iGEM Safety and Security Committee. The Phyllotreta striolata we used in our experiment were collected from Shenzhen university’s field station, which is a very well managed and have strict regulations on biosafety and biosecurity.
The product of our project is siRNA/shRNA that are designed to target the genes of Phyllotreta striolata. When we design the siRNAs, we have BLASTed the siRNA sequences against human genome database to avoid the sequences that have high homologous with the sequences of human genes.