Cre Recombinase Mediates the Removal of Bacterial Backbone to Efficiently Generate rSV40

Molecular Therapy. Methods & Clinical Development
Xiaoxia ShiPiter J Bosma

Abstract

Gene therapy has been shown to be a feasible approach to treat inherited disorders in vivo. Among the currently used viral vector systems, adeno-associated virus (AAV) vectors are the most advanced and have been applied in patients successfully. An important drawback of non-integrating AAV vectors is their loss of expression upon cell division, while repeating systemic administration lacks efficacy due to the induction of neutralizing antibodies. In addition, a significant percentage of the general population is not eligible for AAV-mediated gene therapy due to pre-existing immunity. Development of additional viral vectors may overcome this hurdle. Simian virus 40 (SV40)-derived vectors have been reported to transduce different tissues, including the liver, and prevalence of neutralizing antibodies in the general population is very low. This renders recombinant SV40 (rSV40) vector an interesting candidate for effective (re-)administration. Clinical use of SV40 vectors is in part hampered by less advanced production methods compared to AAVs. To optimize the production of rSV40 and make it better suitable for clinical practice, we developed a production system that relies on Cre recombinase-mediated removal of the bacterial plasm...Continue Reading

Citations

Sep 10, 2019·Mass Spectrometry Reviews·Ana Margarida RodriguesCarla António

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Methods Mentioned

BETA
transfection
PCR
fluorescence-activated
ELISA
Assay
flow cytometry

Software Mentioned

GraphPad Prism

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