For the first time in iGEM competition, our team constructed siRNA for mouse GATA3 GATA3-siRNA-3 (BBa_K3756003) and siRNA for mouse TSLP TSLP-siRNA-2 (BBa_K3756006) and conducted a series of experiments to prove their outstanding gene silencing ability in vitro and in vivo. Details can be found in Results
Verification of parts in vitro
for GATA3-siRNA-3 (BBa_K3756003)
We constructed DNA circuits expressing GATA3-siRNA-3 (BBa_K3756003). We compare the features of our selected parts with other siRNAs for GATA3, namely GATA3-siRNA-1 (BBa_K3756001)and GATA3-siRNA-2 (BBa_K3756002), in mRNA silencing ability and protein inhibition ability. The results shown in Fig.1 show that GATA3-siRNA-3 (BBa_K3756003) outperforms the other two parts GATA3-siRNA-1 (BBa_K3756001)and GATA3-siRNA-2 (BBa_K3756002).
for TSLP-siRNA-2 (BBa_K3756006)
Similarly, We constructed DNA circuits expressing TSLP-siRNA-2 (BBa_K3756006). We compare the features of our selected parts with other siRNAs for TSLP, namely TSLP-siRNA-1 (BBa_K3756005)and GATA3-siRNA-3 (BBa_K3756007), in mRNA silencing ability. The results shown in Fig.2 show that TSLP-siRNA-2 (BBa_K3756006) outperforms the other two parts TSLP-siRNA-1 (BBa_K3756005)and TSLP-siRNA-3 (BBa_K3756007).
Verification of parts in vivo
We administrated both GATA3-siRNA-3 (BBa_K3756003) and TSLP-siRNA-2 (BBa_K3756006) into mice to prove their silencing ability in vivo. The results is shown in Fig.3, and we can see amazing inhibition effect of our two parts on target gene in both mRNA expression and protein expression.
In conclusion, the two parts we designed this year has outstanding inhibition ability on target genes, and we have proved that both in vitro and in vivo experiments, which is sure to benefits other iGEMers or synthetic biology in the future.