Martin Ryan: The Discovery of the 2A Protein Co-expression System and its Utility in Biomedical and Biotechnological Applications – March 15, 2018
Prof. Martin Ryan: Biomedical Sciences Research Complex (BSRC), School of Biology, University of St. Andrews, North Haugh, St. Andrews KY16 9ST. Fife, Scotland, Uk.
e-mail : firstname.lastname@example.org
web page : https://www.fmdv.ac.uk/
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.
Il Professor Martin Ryan sarà ospite del Professor Roberto Giovannoni.
La partecipazione al seminario è libera ed aperta a tutti gli interessati.
Written by Gessica Sala