0000000000307875
AUTHOR
O Argyros
DEVELOPMENT OF S/MAR MINICIRLES VECTOR FOR PERSISTENT EXPRESSION IN VIVO.
An ideal vector for gene therapy must fulfil the following requirements: non-toxicity, mitotic stability and persistent therapeutic levels of transgene expression. Viral vectors are widely used due to their ability to sustain prolonged expression. Their potentially tumorigenic effects are a limiting factor for in vivo applications. Non-viral vectors, which can be designed to be free from viral sequences, are a promising alternative for gene transfer although they often produce transient transgene expression. This limitation of this vector type is primarily due to bacterial sequences they contain and these have proven to be toxic for the mammalian cells as they contain a high number of unmet…
Un vettore NON-virale contenente un elemento S-MAR (Scafold/Matrix Attachment)consente un'espressione persistente nel tessuto epatico nel modello murino.
Un vettore ideale dovrebbe consentire l'espressione di un transgene senza alcuna limitazione di sicurezza e di riproducibilità. Qui riportiamo lo sviluppo di un nuovo vettore NON-virale basato su DNA episomale plasmidico (pDNA) che sembra soddisfare al pieno le caratteristiche del vettore ideale nel tessuto epatico. Questo pDNA deriva dalla combinazione tra un promotore epatospecifico (AAT promoter) posto a monte del transgene e un elemento S-MAR (Scafold/Matrix Attachment) posto a valle, mentre il reporter è gene della luciferasi. L'applicazione nel tessuto epatico è stata effettuata mediante iniezione ad alta pressione per via vena caudale del modello murino (hydrodynamic delivery). L'esp…
Development of S/MAR plasmid vector for persistent expression and maintenance in vivo
An ideal gene therapy vector should enable persistent transgene expression without limitations of safety and reproducibility. Here we report the development of a non-viral episomal plasmid DNA (pDNA) vector that appears to fulfil these criteria. This pDNA vector combines a scaffold/matrix attachment region (S/MAR) with a human liver-specific promoter (a1-antitrypsin (AAT)) in such a way that long-term expression is enabled in murine liver following hydrodynamic injection. Long-term expression is demonstrated by monitoring the longitudinal luciferase expression profile for up to 6 months by means of in situ bioluminescent imaging. We conclude that the combination of a mammalian, tissue-speci…
Development of a new S/MAR containing Minicircle DNA vector: a new promise for Gene Therapy.
A barrier limiting the use of nonviral vectors for gene therapy is related to the short duration of transgene expression in vivo. Development, evaluation, and optimisation of a long term transgene expression using a non viral vector is currently the primary aim in our research field. Recently, we have demonstrate that a nonviral episomal plasmid (pDNA) vector combined with a scaffold/matrix attachment region (S/MAR) is able to sustain long-term expression in murine liver for at least six months following hydrodynamic injection. However, plasmids contain sequences, which are essential for propagation in bacteria but are unnecessary for expression in mammalian cells. These bacterial component…