Characterization of AAV vector particle stability at the single-capsid level

Autor: Julien Bernaud, Cendrine Faivre-Moskalenko, Ludovic Aillot, Lara Gardette, Martin Castelnovo, Axel Rossi, Hildegard Büning, Anny Fis, Anna Salvetti
Přispěvatelé: Laboratoire de Physique des Matériaux (LPM), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon, NUCLEOVIR, Centre de génétique et de physiologie moléculaire et cellulaire (CGPhiMC), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Centre International de Recherche en Infectiologie (CIRI), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure de Lyon (ENS de Lyon)-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Equipe virus AAV et vecteurs AAV (INSERM U758/ ENS de Lyon), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de la Santé et de la Recherche Médicale (INSERM), Hannover Medical School [Hannover] (MHH)
Jazyk: angličtina
Rok vydání: 2018
Předmět:
Zdroj: Journal of Biological Physics
Journal of Biological Physics, 2018, 44 (2), pp.181-194. ⟨10.1007/s10867-018-9488-5⟩
ISSN: 0092-0606
1573-0689
DOI: 10.1007/s10867-018-9488-5⟩
Popis: Virus families have evolved different strategies for genome uncoating, which are also followed by recombinant vectors. Vectors derived from adeno-associated viruses (AAV) are considered as leading delivery tools for in vivo gene transfer, and in particular gene therapy. Using a combination of atomic force microscopy (AFM), biochemical experiments, and physical modeling, we investigated here the physical properties and stability of AAV vector particles. We first compared the morphological properties of AAV vectors derived from two different serotypes (AAV8 and AAV9). Furthermore, we triggered ssDNA uncoating by incubating vector particles to increasing controlled temperatures. Our analyses, performed at the single-particle level, indicate that genome release can occur in vitro via two alternative pathways: either the capsid remains intact and ejects linearly the ssDNA molecule, or the capsid is ruptured, leaving ssDNA in a compact entangled conformation. The analysis of the length distributions of ejected genomes further revealed a two-step ejection behavior. We propose a kinetic model aimed at quantitatively describing the evolution of capsids and genomes along the different pathways, as a function of time and temperature. This model allows quantifying the relative stability of AAV8 and AAV9 particles. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10867-018-9488-5) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
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