Characterization of AAV vector particle stability at the single-capsid level.
J Biol Phys
; 44(2): 181-194, 2018 06.
Article
en En
| MEDLINE
| ID: mdl-29656365
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.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Cápside
/
Dependovirus
/
Vectores Genéticos
Idioma:
En
Revista:
J Biol Phys
Año:
2018
Tipo del documento:
Article
País de afiliación:
Francia