Correction of multi-gene deficiency in vivo using a single 'self-cleaving' 2A peptide-based retroviral vector - PubMed (original) (raw)

doi: 10.1038/nbt957. Epub 2004 Apr 4.

PMID: 15064769
DOI: 10.1038/nbt957

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

Andrea L Szymczak et al. Nat Biotechnol. 2004 May.

Erratum in

Nat Biotechnol. 2004 Dec;22(12):1590
Nat Biotechnol. 2004 Jun;22(6):760

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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Correction of multi-gene deficiency in vivo using a single 'self-cleaving' 2A peptide-based retroviral vector - PubMed (original) (raw)