Translational efficiency of EMCV IRES in bicistronic vectors is dependent upon IRES sequence and gene location
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.
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BibTeX
@article{bochkov2026translational,
title = {{Translational efficiency of EMCV IRES in bicistronic vectors is dependent upon IRES sequence and gene location}},
author = {Yury A Bochkov and A. C. Palmenberg},
journal = {Biotechniques},
year = {2026},
month = oct,
doi = {10.2144/000112243},
url = {https://hal.science/hal-05787854v1},
note = {CC BY},
}
RIS
TY - JOUR
TI - Translational efficiency of EMCV IRES in bicistronic vectors is dependent upon IRES sequence and gene location
AU - Yury A Bochkov
AU - A. C. Palmenberg
PY - 2026
DA - 2026/10/09
JO - Biotechniques
DO - 10.2144/000112243
UR - https://hal.science/hal-05787854v1
AB - 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.
SN - 0736-6205
SN - 1940-9818
ER -