High translation efficiency is mediated by the encephalomyocarditis virus internal ribosomal entry sites if the natural sequence surrounding the eleventh AUG is retained.
ABSTRACT Internal ribosomal entry sites (IRES) allow cap-independent translation and are sequences that are often used in gene therapy for strategies in which several genes need to be expressed. In this study, two different sequences of encephalomyocarditis virus (EMCV) IRES were compared for their translation efficiency in the context of retroviral vectors. When the sequence surrounding the 11th AUG of the IRES was conserved (IRESg), the translation efficiency was significantly higher than if the AUG of the downstream gene started with the 11th AUG of the IRES (IRESb). The translation efficiency with IRESg was influenced by the cell type and also by the nature of the transgene.
Cell cycle (Georgetown, Tex.) 07/2008; 7(12). · 5.36 Impact Factor
Article: Translational efficiency of EMCV IRES in bicistronic vectors is dependent upon IRES sequence and gene location.[show abstract] [hide abstract]
ABSTRACT: The internal ribosomal entry site (IRES)from encephalomyocarditis virus (EMCV) is a popular RNA element used widely in experimental and pharmaceutical applications to express proteins in eukaryotic cells or cell-free extracts. Inclusion of the wild-type element in monocistronic or bicistronic messenger RNAs (mRNAs) confers a high level of cap-independent translation activity to appropriately configured cistrons. The history of this element and the experimental consequences of sequence derivations inherent to commercial IRES vectors are less well known. Compared head-to-head with dual-luciferase reporter constructs, a native EMCV IRES in a bicistronic configuration directed 8- to 10-fold more protein than a similarly configured pIRES vector. It also produced nearly twice as much protein as pCITE-1, an early monocistronic iteration, harboring a suboptimal A7 sequence in a crucial structural motif The results indicate that investigators should be aware of and carefully report the sequence of their IRES in any comparative study. The preferred IRES (viral bases 273-845) and the minimum IRES (viral bases 400-836) for optimum activity are illustrated.BioTechniques 10/2006; 41(3):283-4, 286, 288 passim. · 2.67 Impact Factor
Dataset: Cancer Res 07 Bourbeau