OBJECTIVES The excess proliferation of vascular smooth muscle cells (VSMCs) and the development of intimal hyperplasia is a hallmark of vein graft failure. This study aimed to verify that a single intraoperative transfection of early growth response gene-1 (Egr-1) decoy oligonucleotide (ODN) can suppress vein graft proliferation of VSMCs and intimal hyperplasia. METHODS In a rabbit model, jug...

We have developed an efficient and reproducible method for RNA transfection, using a synthetic cationic lipid, N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA), incorporated into a liposome (lipofectin). Transfection of 10 ng to 5 micrograms of Photinus pyralis luciferase mRNA synthesized in vitro into NIH 3T3 mouse cells yields a linear response of luciferase activity. The...

BACKGROUND Up to now, the different uptake pathways and the subsequent intracellular trafficking of plasmid DNA have been largely explored. By contrast, the mode of internalization and the intracellular routing of an exogenous mRNA in transfected cells are poorly investigated and remain to be elucidated. The bioavailability of internalized mRNA depends on its intracellular routing and its poten...

Abstract: Coxsackievirus B3 (CVB3) is the most common agent known to cause viral myocarditis. The viral genome encodes a single polyprotein that is cleaved to produce several proteins by virally encoded proteases. Most of this proteolytic processing is catalyzed by a cysteine protease called 3C. The 3C protease plays major role in viral replication and cellular damage. To understand the mecha...

When working in the field of target validation, or when studying functional assays requiring the transfection of small interfering RNA (siRNA) into a cell, the presence of having a fast, easy-to-use method to optimize transfection conditions for given cell lines would be of great value. Three to four siRNA sequences are generally tested for each gene in order to suppress the expression of a pro...

background: baculovirus expression system is one of the most attractive and powerful eukaryotic expression systems for the production of recombinant proteins. the presence of a biomarker is required to monitor transfection efficiency or protein expression levels in insect cells. methods: the aim of this study was to construct a baculovirus expression vector encoding a copepod super green fluore...

Background: Germline mutations in the mismatch repair (MMR) genes hMLH1 and hMSH2 can cause hereditary non-polyposis colorectal cancer (HNPCC). However, the functional in vitro analysis of hMLH1 and hMSH2 mutations remains difficult. Aims: To establish an in vitro method for the functional characterisation of hMLH1 and hMSH2 mutations. Methods: hMLH1 and hMSH2 wild type (wt) genes and several m...