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

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 vect...

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Bibliographic Details
Published in:	Nature biotechnology Vol. 22; no. 5; pp. 589 - 594
Main Authors:	Vignali, Dario A A, Szymczak, Andrea L, Workman, Creg J, Wang, Yao, Vignali, Kate M, Dilioglou, Smaroula, Vanin, Elio F
Format:	Journal Article
Language:	English
Published:	New York, NY Nature 01-05-2004 Nature Publishing Group
Subjects:	Amino Acid Sequence Animals Biological and medical sciences Biotechnology Blood Bone marrow Cell Line Energy transfer Fluorescence Fundamental and applied biological sciences. Psychology Genetic Vectors Humans Mice Mice, Inbred C57BL Molecular Sequence Data Peptides Peptides - chemistry Peptides - genetics Peptides - metabolism Proteins Resonance Retroviridae - genetics Sequence Homology, Amino Acid Stem cells Spleen T cell receptor Peptides Rodentia Fluorescence Retroviridae Resonance energy Complexes Genetic translation Protein Virus In vivo Vertebrata Blood cell Mammalia Gene Mouse Production Bone marrow Cleavage Gene therapy Energy transfer Vector
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Summary:	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 (CD3 , γ, δ, ζ), 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.
Bibliography:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 SourceType-Other Sources-1 content type line 63 ObjectType-Correspondence-1
ISSN:	1087-0156 1546-1696
DOI:	10.1038/nbt957