Figure 1.

Silencer and reporter constructs used to test RNAi efficacy. (A) The silencer construct, pSi, includes two cassettes for easy exchange of promoters and shRNAs with or without Ago2 co-expression. The shRNA promoter fragments can be inserted into pSi through the SacI site on the vector and the shRNA fragments can then be inserted into pSi at XhoI and EcoRI sites. (B) The reporter construct, pRe, contains one CMV or NSE promoter for GFP expression and another CMV promoter for mCherry expression. The GFP silencing effect by shRNAs can be directly evaluated by calculating the green to red fluorescence intensity ratio in individual cells.