Production, purification, crystallization and preliminary X-ray analysis of adeno-associated virus serotype 8.
Adeno-associated viruses (AAVs) are actively being developed for clinical gene-therapy applications and the efficiencies of the vectors could be significantly improved by a detailed understanding of their viral capsid structures and the structural determinants of their tissue-transduction interactions. AAV8 is approximately 80% identical to the more widely studied AAV2, but its liver-transduction efficiency is significantly greater than that of AAV2 and other serotypes. The production, purification, crystallization and preliminary X-ray crystallographic analysis of AAV8 viral capsids are reported. The crystals diffract X-rays to 3.0 A resolution using synchrotron radiation and belong to the hexagonal space group P6(3)22, with unit-cell parameters a = 257.5, c = 443.5 A. The unit cell contains two viral particles, with ten capsid viral protein monomers per crystallographic asymmetric unit.
Lane
Michael Douglas
MD
Department of Biochemistry and Molecular Biology, Center for Structural Biology, McKnight Brain Institute, University of Florida, Gainesville, FL 32610, USA.
Nam
Hyun Joo
HJ
Padron
Eric
E
Gurda-Whitaker
Brittney
B
Kohlbrenner
Eric
E
Aslanidi
George
G
Byrne
Barry
B
McKenna
Robert
R
Muzyczka
Nicholas
N
Zolotukhin
Sergei
S
Agbandje-McKenna
Mavis
M
D000818:Animals; D002213:Capsid / Q000737:chemistry*; D036022:Capsid Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D003460:Crystallization / Q000379:methods; D000229:Dependovirus / Q000737:chemistry*; D007313:Insecta; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D014961:X-Ray Diffraction
D036022:Capsid Proteins; D014764:Viral Proteins