US 12,188,037 B2
Synthetic combinatorial AAV3 capsid library
Sergei Zolotukhin, Gainesville, FL (US); and Damien Marsic, Rockville, MD (US)
Assigned to University of Florida Research Foundation, Incorporated, Gainesville, FL (US)
Appl. No. 15/769,615
Filed by University of Florida Research Foundation, Incorporated, Gainesville, FL (US)
PCT Filed Oct. 21, 2016, PCT No. PCT/US2016/058130
§ 371(c)(1), (2) Date Apr. 19, 2018,
PCT Pub. No. WO2017/070476, PCT Pub. Date Apr. 27, 2017.
Claims priority of provisional application 62/245,025, filed on Oct. 22, 2015.
Prior Publication US 2020/0181644 A1, Jun. 11, 2020
Int. Cl. C12N 15/86 (2006.01); C07K 14/005 (2006.01); C12N 7/00 (2006.01); C40B 40/08 (2006.01)
CPC C12N 15/86 (2013.01) [C07K 14/005 (2013.01); C12N 7/00 (2013.01); C40B 40/08 (2013.01); C12N 2750/14122 (2013.01); C12N 2750/14143 (2013.01); C12N 2810/6027 (2013.01)] 4 Claims
 
1. A non-naturally occurring nucleic acid molecule comprising:
(a) a first nucleic acid sequence encoding at least one AAV Rep protein from serotype 3; and
(b) a second nucleic acid sequence encoding an AAV3B VP1 Cap protein;
wherein the AAV3B VP1 Cap protein comprises a variant variable region VII (VR-VII) comprising a plurality of amino acid substitutions relative to the wildtype AAV3B VP1 Cap protein VR-VII sequence set forth in amino acids 306-317 of SEQ ID NO: 4, wherein the variant VR-VII comprises:
a first amino acid that corresponds to amino acid 306 of SEQ ID NO: 4 and is E;
a second amino acid that corresponds to amino acid 307 of SEQ ID NO: 4 and is independently selected from the group consisting of N, S and D;
a third amino acid that corresponds to amino acid 308 of SEQ ID NO: 4 and is independently selected from the group consisting of S, G and A;
a fourth amino acid that corresponds to amino acid 309 of SEQ ID NO: 4 and is independently selected from the group consisting of T, K, N, R, S, E, D and A;
a fifth amino acid that corresponds to amino acid 310 of SEQ ID NO: 4 and is independently selected from the group consisting of K, T, R, E and G;
a sixth amino acid that corresponds to amino acid 311 of SEQ ID NO: 4 and is independently selected from the group consisting of N, T, D, A and G;
a seventh amino acid that corresponds to amino acid 312 of SEQ ID NO: 4 and is D;
an eighth amino acid that corresponds to amino acid 313 of SEQ ID NO: 4 and is independently selected from the group consisting of A, T and I;
a ninth amino acid that corresponds to amino acid 314 of SEQ ID NO: 4 and is independently selected from the group consisting of E and A;
a tenth amino acid that corresponds to amino acid 315 of SEQ ID NO: 4 and is independently selected from the group consisting of N, I, D, V, Y and F;
an eleventh amino acid that corresponds to amino acid 316 of SEQ ID NO: 4 and is independently selected from the group consisting of E and G; and
a twelfth amino acid that corresponds to amino acid 317 of SEQ ID NO: 4 and is independently selected from the group consisting of K, R, S, Q, H, E, D and G.