Esophageal Adenocarcinoma Methods and Protocols

Fig. 1 Delivery and mechanism of RNAi via shRNA lentiviral particles in esophageal adenocarcinoma cells.
Lentiviral particles deliver target shRNA (MET shRNA) to the target cells of esophageal adenocarcinoma. The
shRNA then integrate to the genomic DNA of cells followed by reverse-transcription. The integrated shRNA is
then transcribed by polymerase III, resulting in pri-microRNA. This pri-microRNA is processed by a multi-
enzyme complex Drosha to generate pre-shRNA. Pre-shRNA is then exported to the cytoplasm and cleavage
by Dicer complex to produce siRNA. The double-stranded siRNA unwind into two single-stranded small RNA
(passenger strand and the guide strand). After that, the passenger strand degraded. The guide RNA strand
loaded on RNA-induced silencing complex (RISC), which locates the complementary sequence and induce
molecular cleavage of target MET mRNA by a catalytic component of RISC, Argonaute 2 (Ago2). The cleaved
mRNA is degraded rapidly thereby template for protein production is not available for the target genes