Figure 1. (1) The lentiviral particle binds to the host cell and delivers its engineered RNA genome, which includes encoded microRNA, to the cytoplasm. (2) The viral genome is reverse-transcribed in the cytoplasm. The DNA intermediate form is imported into the nucleus and is stably integrated into the host genome.
(3) shMIMIC microRNA transgenes are transcribed in the same manner as endogenous microRNA genes. Native and over-expressed primary microRNAs are processed by the Drosha complex to yield the precursor microRNA (4), which is then shuttled out of the nucleus into the cytoplasm (5) and further processed by the Dicer complex into double-stranded microRNA (6). One strand is selected for incorporation into the RNA-induced Silencing Complex (RISC). microRNA-programmed RISC binds to target mRNA causing transcript destabilization and subsequent down-regulation of protein expression (7).
Figure 2. TurboGFP expression in four different cell lines (HEK293, HUVEC, K562, SH-SY5Y) visualized by epifluorescence microscopy 120 hours post-transduction with various shMIMIC microRNA (no puromycin selection).
Figure 3. HEK293, HUVEC, K562, or SH-SY5Y cells were transduced in biological triplicate with specific shMIMIC microRNA or negative control viral particles.