In this work, we investigated the potential role of the small G protein RhoA in ethanol-induced tight junction (TJ) protein disassembly and increased intestinal epithelial barrier (IEB) permeability. Our study used Caco-2 cells as an in vitro IEB model and RhoA short hairpin RNA (shRNA) interference to establish whether RhoA plays a role in ethanol-induced TJ opening. RhoA shRNA interference pa...

Use of RNA interference (RNAi) in forward genetic screens is proliferating. Currently, short-interfering RNAs (siRNAs) and short-hairpin RNAs (shRNAs) are being used to silence genes to tease out functional information. It is becoming easier to harness RNAi to silence specific genes, owing to the development of libraries of readymade shRNA and siRNA gene-silencing constructs by using a variety ...

RNA interference (RNAi) mediated by siRNAs has proved to be a highly effective gene silencing mechanism with great potential for gene therapeutic applications. However, siRNA agents have been shown to exert non-target-related biological effects and toxicities, including immune stimulation. Specifically, siRNA synthesized from a T7 RNA polymerase system can trigger the potent induction of type I...

Short hairpin RNAs (shRNAs) are effective in generating stable repression of gene expression. RNA polymerase III (RNAP III) type III promoters (U6 or H1) are typically used to drive shRNA expression. While useful for some knockdown applications, the robust expression of U6/H1-driven shRNAs can induce toxicity and generate heterogeneous small RNAs with undesirable off-target effects. Additionall...

BACKGROUND Short hairpin RNA (shRNA) encoded within an expression vector has proven an effective means of harnessing the RNA interference (RNAi) pathway in mammalian cells. A survey of the literature revealed that shRNA vector construction can be hindered by high mutation rates and the ensuing sequencing is often problematic. Current options for constructing shRNA vectors include the use of ann...

BACKGROUND Coxsackievirus B3 is an important infectious agent of viral myocarditis, pancreatitis and aseptic meningitis, but there are no specific antiviral therapeutic reagents in clinical use. RNA interference-based technology has been developed to prevent the viral infection. METHODS To evaluate the impact of RNA interference on viral replication, cytopathogenicity and animal survival, sho...

RNA interference (RNAi) is widely used to study gene functions as a reverse genetic means from first-generation siRNA to second-generation short hairpin RNA (shRNA) or the newly developed microRNA (shRNA-miR). Here we report a gene knockdown vector system based on the mouse miR-21 hairpin structure. In this system, the pre-miRNA hairpin of the miR-21 gene was modified by replacing the 22-nucleo...

in vitro, specifically decreasing GLI1 messenger RNA (mRNA) levels, at concentrations ranging from 1 to 10mM showing dose-dependent effects, modulated by serum levels. The best tests for specificity rely on mimicry by targeting SMOH with RNA interference (RNAi), the use of insensitive SMOH mutants and pathway epistatic analyses. Targeting SMOH through lentivector-mediated short hairpin RNA (shR...

Pancreatic cancer is known for its notorious fast progression and poor prognosis. Long noncoding RNA (lncRNA) AL161431.1 has been reported to be involved in the pathogenesis of different cancers. In this study, we explored role lncRNA development pancreatic by bioinformatic analysis, vitro vivo experiments BxPC-3 SW1990 cells, as well clinical samples. We found that was highly expressed cells t...