8A9U

Full AAV3B-VP1KO virion

Summary for 8A9U

• Entry DOI: 10.2210/pdb8a9u/pdb
• EMDB information: 15286
• Descriptor: Capsid protein VP1 (1 entity in total)
• Functional Keywords: adeno-associated virus, aav, aav3b, aav serotype 3b, full, vp1ko, virus
• Biological source: Homo sapiens (human)
• Total number of polymer chains: 1
• Total formula weight: 66728.28

Authors

Arriaga, I.,Abrescia, N.G.A. (deposition date: 2022-06-29, release date: 2022-09-21, Last modification date: 2024-11-13)

Primary citation

Arriaga, I.,Navarro, A.,Etxabe, A.,Trigueros, C.,Samulski, R.J.,Moullier, P.,Francois, A.,Abrescia, N.G.A.
Cellular and Structural Characterization of VP1 and VP2 Knockout Mutants of AAV3B Serotype and Implications for AAV Manufacturing.
Hum Gene Ther, 33:1142-1156, 2022

PubMed Abstract: AAV virion biology is still lacking a complete understanding of the role that the various structural subunits (VP1, 2, and 3) play in virus assembly, infectivity, and therapeutic delivery for clinical indications. In this study, we focus on the less studied adeno-associated virus AAV3B and generate a collection of AAV plasmid substrates that assemble virion particles deficient specifically in VP1, VP2, or VP1 and 2 structural subunits. Using a collection of biological and structural assays, we observed that virions devoid of VP1, VP2, or VP1 and 2 efficiently assembled virion particles, indistinguishable by cryoelectron microscopy (cryo-EM) from that of wild type (WT), but unique in virion transduction (WT > VP2 > VP1 > VP1 and 2 mutants). We also observed that the missing structural subunit was mostly compensated by additional VP3 protomers in the formed virion particle. Using cryo-EM analysis, virions fell into three classes, namely full, empty, and partially filled, based on comparison of density values within the capsid. Further, we characterize virions described as "broken" or "disassembled" particles, and provide structural information that supports the particle dissolution occurring through the two-fold symmetry sites. Finally, we highlight the unique value of employing cryo-EM as an essential tool for release criteria with respect to AAV manufacturing.

PubMed: 36082996
DOI: 10.1089/hum.2022.119

Experimental method

ELECTRON MICROSCOPY (3.1 Å)