TY - JOUR
T1 - Atomic structure of a rationally engineered gene delivery vector, AAV2.5
AU - Burg, Matthew
AU - Rosebrough, Claire
AU - Drouin, Lauren M.
AU - Bennett, Antonette
AU - Mietzsch, Mario
AU - Chipman, Paul
AU - McKenna, Robert
AU - Sousa, Duncan
AU - Potter, Mark
AU - Byrne, Barry
AU - Jude Samulski, R.
AU - Agbandje-McKenna, Mavis
N1 - Funding Information:
We thank the University of Florida (UF) Interdisciplinary Center for Biotechnology Research (ICBR) electron microscopy lab for providing negative-stain electron microscope services and the UF Division of Sponsored Programs and college of Medicine for providing funds for the purchase of the FEI Spirit and TF20 microscopes. Data collection at Florida State University was made possible by NIH grants S10 OD018142-01 Purchase of a direct electron camera for the Titan-Krios at FSU (PI Taylor), S10 RR025080-01 Purchase of a FEI Titan Krios for 3-D EM (PI Taylor), and U24 GM116788 The Southeastern Consortium for Microscopy of MacroMolecular Machines (PI Taylor). We thank Olga G. Kozyreva for generating the expression plasmid for AAV2.5. This work was supported by NIH R01 AI072176 to RJS and MAM and NIH R01 GM082946 to RM and MAM.
Publisher Copyright:
© 2018
PY - 2018/9
Y1 - 2018/9
N2 - AAV2.5 represents the first structure-guided in-silico designed Adeno-associated virus (AAV) gene delivery vector. This engineered vector combined the receptor attachment properties of AAV serotype 2 (AAV2) with the muscle tropic properties of AAV1, and exhibited an antibody escape phenotype because of a modified antigenic epitope. To confirm the design, the structure of the vector was determined to a resolution of 2.78 Å using cryo-electron microscopy and image reconstruction. The structure of the major viral protein (VP), VP3, was ordered from residue 219 to 736, as reported for other AAV structures, and the five AAV2.5 residues exchanged from AAV2 to AAV1, Q263A, T265 (insertion), N706A, V709A, and T717N, were readily interpretable. Significantly, the surface loops containing these residues adopt the AAV1 conformation indicating the importance of amino acid residues in dictating VP structure.
AB - AAV2.5 represents the first structure-guided in-silico designed Adeno-associated virus (AAV) gene delivery vector. This engineered vector combined the receptor attachment properties of AAV serotype 2 (AAV2) with the muscle tropic properties of AAV1, and exhibited an antibody escape phenotype because of a modified antigenic epitope. To confirm the design, the structure of the vector was determined to a resolution of 2.78 Å using cryo-electron microscopy and image reconstruction. The structure of the major viral protein (VP), VP3, was ordered from residue 219 to 736, as reported for other AAV structures, and the five AAV2.5 residues exchanged from AAV2 to AAV1, Q263A, T265 (insertion), N706A, V709A, and T717N, were readily interpretable. Significantly, the surface loops containing these residues adopt the AAV1 conformation indicating the importance of amino acid residues in dictating VP structure.
KW - Adeno-associated virus
KW - Parvovirus
KW - Structure-guided design
KW - Vector engineering
KW - cryo-EM
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U2 - 10.1016/j.jsb.2018.05.004
DO - 10.1016/j.jsb.2018.05.004
M3 - Article
C2 - 29775653
AN - SCOPUS:85047059161
SN - 1047-8477
VL - 203
SP - 236
EP - 241
JO - Journal of Structural Biology
JF - Journal of Structural Biology
IS - 3
ER -