Versatile co-expression of graft-protective proteins using 2A-linked cassettes
Nella Fisicaro, Sarah L Londrigan, Jamie L Brady, Evelyn Salvaris, Mark B Nottle, Philip J O'Connell, Simon C Robson, Anthony JF d'Apice, Andrew M Lew, Peter J Cowan
Xenotransplantation | WILEY | Published : 2011
BACKGROUND: Expression of multiple graft-protective proteins targeted to different locations (i.e., intracellular, cell surface, and secreted) has become an increasingly important goal in xenotransplantation. The 2A "ribosome skip" signal is used as a linker to enable transgene co-expression, but some studies have shown that post-translational modification and trafficking of 2A-linked proteins may be adversely affected depending on their position relative to 2A. We tested whether several relevant proteins, subject to a range of processing and localization mechanisms, could be efficiently co-expressed using the 2A system. METHODS: Six expression cassettes were constructed, each containing up ..View full abstract
Awarded by National Institutes of Health USA
Awarded by NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
This work was supported by the Australian National Health and Medical Research Council, the National Institutes of Health USA (to SCR and AJFD; U01 AI066331 and P01AI045897), and the Juvenile Diabetes Research Foundation. We thank Helen Barlow and the staff of the Bioresources Centre, St Vincent's Hospital Melbourne, for the generation and care of transgenic mice.