Skip to Main content Skip to Navigation
New interface
Journal articles

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

Julien Bernaud 1 Axel Rossi 2 Anny Fis 1 Lara Gardette 1 Ludovic Aillot 2 Hildegard Büning 3 Martin Castelnovo 1 Anna Salvetti 2 Cendrine Faivre-Moskalenko 1 
CGPhiMC - Centre de génétique et de physiologie moléculaire et cellulaire, CIRI - Centre International de Recherche en Infectiologie - UMR, INSERM U758/ ENS de Lyon - Equipe virus AAV et vecteurs AAV
Abstract : 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.
Complete list of metadata
Contributor : Christelle Cheval Connect in order to contact the contributor
Submitted on : Tuesday, January 8, 2019 - 4:42:11 PM
Last modification on : Saturday, September 24, 2022 - 3:10:05 PM

Links full text




Julien Bernaud, Axel Rossi, Anny Fis, Lara Gardette, Ludovic Aillot, et al.. Characterization of AAV vector particle stability at the single-capsid level. Journal of Biological Physics, 2018, 44 (2), pp.181-194. ⟨10.1007/s10867-018-9488-5⟩. ⟨hal-01974270⟩



Record views