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Genetics. 2014 Mar;196(3):605-13. doi: 10.1534/genetics.113.160846. Epub 2013 Dec 20.

Simultaneous expression of multiple proteins under a single promoter in Caenorhabditis elegans via a versatile 2A-based toolkit.

Author information

1
Department of Development and Stem Cells, Institut de Génétique et de Biologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique Unité Mixte de Recherche 7104/Institut National de la Santé et de la Recherche Médicale U964/Université de Strasbourg, 67404 Illkirch CU Strasbourg, France.

Abstract

Caenorhabditis elegans is a powerful in vivo model in which transgenesis is highly developed. However, while the analysis of biological phenomena often require the expression of more than one protein of interest, no reliable tool exists to ensure efficient concomitant and equivalent expression of more than two polypeptides from a single promoter. We report the use of viral 2A peptides, which trigger a "ribosomal-skip" or "STOP&GO" mechanism during translation, to express multiple proteins from a single vector in C. elegans. Although none of the viruses known to infect C. elegans contain 2A-like sequences, our results show that 2A peptides allow the production of separate functional proteins in all cell types and at all developmental stages tested in the worm. In addition, we constructed a toolkit including a 2A-based polycistronic plasmid and reagents to generate 2A-tagged fosmids. 2A peptides constitute an important tool to ensure the delivery of multiple polypeptides in specific cells, enabling several novel applications such as the reconstitution of multi-subunit complexes.

KEYWORDS:

2A peptide; Caenorhabditis elegans; SL2; internal ribosomal entry site (IRES); stoichiometric delivery of multiple polypeptides

PMID:
24361941
PMCID:
PMC3948794
DOI:
10.1534/genetics.113.160846
[Indexed for MEDLINE]
Free PMC Article

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