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Quantitative comparison of gene co-expression in a bicistronic vector harboring IRES or coding sequence of porcine teschovirus 2A peptide

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Abstract

In biotechnology, simultaneous expression of more than one target gene is often required. Multicistronic vectors encoding several proteins are being actively developed for this purpose. Most often, the commercially available vectors utilize various types of internal ribosomal entry site of the encephalomyocarditis virus (IRES EMCV). However, many researchers consider bicistronic vectors on the basis of sequences that encode self-cleaving 2A peptides more promising. In the work, we compare the efficiency of gene expression in cells transfected with bicistronic constructs bearing either IRES EMCV or the P2A nucleotide sequence corresponding to the porcine teschovirus-1 2A peptide. Efficiency of gene expression was determined in three mammalian cell lines by measurement of co-expression levels of genes coding for RFP and EGFP proteins linked by IRES or P2A sequence. A higher level of the transgene expression was detected in cells transfected with P2A sequence-based genetic constructs.

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Abbreviations

UTR:

untranslated region

RF:

reading frame

EPR:

endoplasmic reticulum

CMV:

cytomegalovirus promoter

EGFP:

green fluorescent protein

IRES:

internal ribosomal entry site

NMD:

non-sense mediated decay

P2A and P2A :

porcine 2A peptide and the coding sequence

RFP:

red fluorescent protein

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Authors and Affiliations

  1. Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russia

    A. I. Kuzmich, A. V. Vvedenskii, E. P. Kopantzev & T. V. Vinogradova

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  1. A. I. Kuzmich
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  2. A. V. Vvedenskii
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  3. E. P. Kopantzev
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  4. T. V. Vinogradova
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Correspondence to A. I. Kuzmich.

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Original Russian Text © A.I. Kuzmich, A.V. Vvedenskii, E.P. Kopantzev, T.V. Vinogradova, 2013, published in Bioorganicheskaya Khimiya, 2013, Vol. 39, No. 4, pp. 454–465.

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Kuzmich, A.I., Vvedenskii, A.V., Kopantzev, E.P. et al. Quantitative comparison of gene co-expression in a bicistronic vector harboring IRES or coding sequence of porcine teschovirus 2A peptide. Russ J Bioorg Chem 39, 406–416 (2013). https://doi.org/10.1134/S1068162013040122

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  • DOI: https://doi.org/10.1134/S1068162013040122

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