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

• Published in: Genetics (Austin) Vol. 196; no. 3; pp. 605 - 613
• Main Authors: Ahier, Arnaud, Jarriault, Sophie
• Format: Journal Article
• Language: English
• Published: United States Genetics Society of America 01.03.2014; Oxford University Press
• Subjects: Animals; Biochemistry, Molecular Biology; Caenorhabditis elegans; Caenorhabditis elegans - embryology; Caenorhabditis elegans - genetics; Development Biology; Gene Expression; Genetic Techniques; Genetic Vectors; Genetics; Investigations; Life Sciences; Peptides - genetics; Peptides - metabolism; Polypeptides; Promoter Regions, Genetic; Proteins; Ribosomes - genetics; Transfection; Viruses - chemistry; SL2; 2A peptide; Caenorhabditis elegans; internal ribosomal entry site (IRES); stoichiometric delivery of multiple polypeptides
• ISSN: 1943-2631; 0016-6731; 1943-2631
• DOI: 10.1534/genetics.113.160846

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.