Alternative-splicing-based bicistronic vectors for ratio-controlled protein expression and application to recombinant antibody production.

Abstract:

:In the last decade polycistronic vectors have become essential tools for both basic science and gene therapy applications. In order to co-express heterologous polypeptides, different systems have been developed from Internal Ribosome Entry Site (IRES) based vectors to the use of the 2A peptide. Unfortunately, these methods are not fully suitable for the efficient and reproducible modulation of the ratio between the proteins of interest. Here we describe a novel bicistronic vector type based on the use of alternative splicing. By modifying the consensus sequence that governs splicing, we demonstrate that the ratio between the synthesized proteins could easily vary from 1 : 10 to 10 : 1. We have established this system with luciferase genes and we extended its application to the production of recombinant monoclonal antibodies. We have shown that these vectors could be used in several typical cell lines with similar efficiencies. We also present an adaptation of these vectors to hybrid alternative splicing/IRES constructs that allow a ratio-controlled expression of proteins of interest in stably transfected cell lines.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Fallot S,Ben Naya R,Hieblot C,Mondon P,Lacazette E,Bouayadi K,Kharrat A,Touriol C,Prats H

doi

10.1093/nar/gkp716

subject

Has Abstract

pub_date

2009-11-01 00:00:00

pages

e134

issue

20

eissn

0305-1048

issn

1362-4962

pii

gkp716

journal_volume

37

pub_type

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