4.5 Article

A bicistronic lentiviral vector based on the 1D/2A sequence of foot-and-mouth disease virus expresses proteins stoichiometrically

Journal

JOURNAL OF BIOTECHNOLOGY
Volume 146, Issue 3, Pages 138-142

Publisher

ELSEVIER
DOI: 10.1016/j.jbiotec.2010.01.017

Keywords

Lentivirus; Foot-and-mouth disease virus; Bicistronic vector; Bicistronic sequence; Internal ribosome entry site; 2A sequence; 2A peptide; Stoichiometric expression; Neurobiology

Funding

  1. La Universidad de Antofagasta (Direccion de Investigacion) [1314, 1315]
  2. El Fondo Nacional de Desarrollo Cientifico y Tecnologico (FONDECYT) [1070462]
  3. NIH [R21MH085954, P01N5053862]
  4. NATIONAL INSTITUTE OF MENTAL HEALTH [R21MH085954] Funding Source: NIH RePORTER
  5. NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE [P01NS053862] Funding Source: NIH RePORTER

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Classic IRES sequences are notorious for exerting biased expression in favor of upstream coding regions when placed into polycistronic vectors. Here, we report the development of a bicistronic lentiviral system based on the 1D/2A sequence from the foot-and-mouth disease virus that is able to maintain tightly balanced control of upstream and downstream protein expression for several days at a stoichiometry very closely approaching 1.0. Our results suggest that the 1D/2A sequence can be optimized in an FUGW lentiviral setting to coordinate expression of multiple polypeptides, presenting a potentially valuable tool to signaling network researchers and to the gene therapy community. (C) 2010 Elsevier B.V. All rights reserved.

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