Characterization of AAVrh.10 as a vector to deliver long-lasting genetic constructs into neurons of neonatal mice following intravenous injection
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Date
2019
Authors
Martin, Matthew
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Abstract
Prion diseases are a fatal neurodegenerative disease caused by the misfolding of the cellular prion protein, PrPC into an infectious isoform, PrPSc. Accumulation of this infectious isoform leads to the damage and death of neurons, and the progression of the disease. The molecular events involved in the pathogenesis of this disease are poorly understood and uncovering them has proven challenging due to the difficulties of identifying and isolating degenerating neurons for analysis. Ultimately, the aim of this project is to characterize the use of AAVrh.10 to deliver genetic constructs into the neurons of mice in vitro, and in vivo. AAVrh.10 was compared to AAV8, and AAV9 to assess transduction efficiencies in primary hippocampal neuron cell cultures. All 3 AAV serotypes were found to transduce primary neurons with high efficiency, and low toxicity. Further work showed that intravenous injection of AAVrh.10 into neonatal mice via the superficial temporal vein was capable of inducing long-lasting transgene expression in neurons and astrocytes within the brain, throughout the entire course of prion disease. IHC analysis is performed to assess the distribution of the AAVrh.10 throughout multiple regions of mouse brain. In addition, the kinetics of the vector concentration in the brain over time will be described. This work is the basis for engineering vectors capable of targeting specific neurons in mouse brain tissue in future studies to understand prion pathogenesis and prion strains in vivo.
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Keywords
Adeno-associated viruses, Prions, Neurodegeneration, Viral Therapy