The Discovery of the 2A Protein Co-expression System and its Utility in Biomedical and Biotechnological Applications

Invited Research Seminar
Giovedì 15 Marzo 2018 Ore 13 Edificio U8-Aula 6
Dipartimento di Medicina e Chirurgia, Via Cadore 48, Monza.

Prof. Martin Ryan
Biomedical Sciences Research Complex (BSRC), School of Biology, University of St. Andrews Fife, Scotland, UK.

"The Discovery of the 2A Protein Co-expression System and its Utility in Biomedical and Biotechnological Applications"

The production of many high-value therapeutic proteins, or, the repair of genetic lesions frequently requires the efficient co-expression of multiple, different, proteins within the same cell.

Transfection of a population of cells using either a mixture of plasmids, or, a single plasmid encoding multiple genes – driven by multiple promoters – only produces a small proportion of transfected cells co-expressing all the genes of interest. 2A and ‘2A-like’ sequences are oligopeptide sequences (18-25aa) which may be used as linker sequences to concatenate multiple genes into a single transgene.

Here, the stop codons of genes are removed and replaced by 2A: a single, long, open reading frame is created – a single transgene driven by a single promoter. When the mRNA is translated, however, the translation product is not a fusion protein: translation stops and then re-starts at each 2A linker sequence – each gene is translated as a separate protein product. In this manner, sequences encoding the constituents of multimeric complexes, biochemical pathways etc. can be co-expressed with high efficiency.

Furthermore, the function of co-translational signal (leader) sequences is not impaired by the 2A system such that individual translation products can be targeted to specific sub-cellular sites. The seminar will describe the discovery and characterisation of these 2A sequences and their use in a wide range of biomedical applications – e.g. production of therapeutic monoclonal antibodies, human gene therapies, the production of human induced pluripotent stem cells (iPSCs) and genome editing technologies.

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Ospite: Dr. Roberto Giovannoni

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