Results Psd-95 Shrna

To address the unsolved question of PSD-95 function I used short hairpin RNA (shRNA) to knock-down protein expression. shRNA mediated knock-down of protein expression is one of the established methods exploiting the naturally occurring expression control mechanism known as RNA interference (RNAi). Cells express shRNAs that are complementary to the mRNA they control. The shRNAs are processed and bound by two complexes called DICER (***) and RISC. Subsequentially the complementary the RISC complex binds the mRNA and the mRNA is degraded. This mRNA depletion decreases the protein level. In shRNA experiments, the short hairpin RNAs are expressed from a DNA plasmid vector. Polymerase III promoters are generally used as they reliably terminate transcription. shRNA constructs affect the targeted mRNA with variable efficiency. I generated four potential shRNA constructs to suppress PSD-95 expression. To test for efficiency I co-expressed the shRNA together with PSD-95 cDNA in HEK293T cells. I used the shRNA vector without a hairpin insert as a control to visualize unaffected PSD-95 expression.