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Correction of multi-gene deficiency in vivo using a single 'self-cleaving' 2A peptide–based retroviral vector

Szymczak, Andrea L, Workman, Creg J, Wang, Yao, Vignali, Kate M, Dilioglou, Smaroula, Vanin, Elio F, Vignali, Dario A A·2004·Nature Biotechnology
DOI 10.1038/nbt957
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Abstract

Attempts to generate reliable and versatile vectors for gene therapy and biomedical research that express multiple genes have met with limited success. Here we used Picornavirus 'self-cleaving' 2A peptides, or 2A-like sequences from other viruses, to generate multicistronic retroviral vectors with efficient translation of four cistrons. Using the T-cell receptor:CD3 complex as a test system, we show that a single 2A peptide-linked retroviral vector can be used to generate all four CD3 proteins (CD3epsilon, gamma, delta, zeta), and restore T-cell development and function in CD3-deficient mice. We also show complete 2A peptide-mediated 'cleavage' and stoichiometric production of two fluorescent proteins using a fluorescence resonance energy transfer-based system in multiple cell types including blood, thymus, spleen, bone marrow and early stem cell progenitors.

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Metadata

Authors
Szymczak, A. L., Workman, C. J., Wang, Y., Vignali, K. M., Dilioglou, S., Vanin, E. F., & Vignali, D. A. A.
Venue
Nature Biotechnology
Publisher
Springer Science and Business Media LLC
Published
2004-04-04
Volume
22
Issue
5
Pages
589-594
ISSN
  • 1087-0156
  • 1546-1696
DOI
10.1038/nbt957
Cited by
1,059
References
1
Language
en
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