Methods and compositions for improving the assembly of adeno-associated viruses (AAVs)
AssigneesMassachusetts Eye and Ear Infirmary, The Schepens Eye Research Institute, Inc.
InventorsAnna C. Maurer, Luc H. Vandenberghe
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Abstract
Described herein are methods and compositions for modifying cells to improve the assembly of adeno-associated viruses (AAVs).
Claims18
A method for improving the assembly of adeno-associated virus (AAV) in a cell, the method comprising:
- increasing expression or activity of Hsc/Hsp70 or a co-factor thereof, vacuolar-specific H+ ATPase, and/or cyclin-dependent kinase 2 in the cell to produce a modified cell;
- infecting the modified cell with an AAV vector to produce an infected modified cell;
- culturing the infected modified cell;
- and
- collecting the assembled AAV.
A method for improving the assembly of adeno-associated virus (AAV), the method comprising:
- producing AAV from a modified cell with the AAV, wherein the cell has been modified to exhibit an increase or decrease in one or more genes or the protein encoded therefrom selected from CHMP7, CEP72, CNOT6, SLC9A6, PPHLN1, ATF7IP, SETDB1, GPR89B, KIF16B, ATAD3A, BAG2, STUB1, DNAJA1, DNAJC7, HSPA8, HSPAIA, CDK1, CDK2, CHCHD2, C1QBP, DPYSL5, FXR1, FXR2, IPO5, LBR, MTPN, NPM3, NPM1, PPL, SNX3, UBE2O, SAE1, CCDC124, GNB1, RABIA, GNB4, RPL23, CKB, SRP9, UCHL1, and TBCB;
- and
- collecting the assembled AAV.
A cell-free culture system for assembling adeno-associated virus comprising:
- culture media;
- and
- at least two proteins, or nucleic acids encoding the at least two proteins, selected from CHMP7, CEP72, CNOT6, SLC9A6, PPHLN1, ATF7IP, SETDB1, GPR89B, KIF16B, ATAD3A, BAG2, STUB1, DNAJA1, DNAJC7, HSPA8, HSPAIA, CDK1, CDK2, CHCHD2, C1QBP, DPYSL5, FXR1, FXR2, IPO5, LBR, MTPN, NPM3, NPM1, PPL, SNX3, UBE2O, SAE1, CCDC124, GNB1, RABIA, GNB4, RPL23, CKB, SRP9, UCHL1, and TBCB.
A cell line comprising:
- one or more mutations in Hsc/Hsp70 or a co-factor thereof, vacuolar-specific H+ ATPase, and/or cyclin-dependent kinase 2;
- and/or
- one or more exogenous constructs expressing Hsc/Hsp70 or a co-factor thereof, vacuolar-specific H+ ATPase, and/or cyclin-dependent kinase 2.
An article of manufacture for improving the assembly of adeno-associated virus (AAV) in a cell, comprising at least one member from at least two of (a), (b) and of (c):
- (a) Hsc/Hsp70 or a co-factor thereof, a nucleic acid encoding Hsc/Hsp70 or a co-factor thereof, or a compound that modulates Hsc/Hsp70 or a co-factor thereof;
- (b) vacuolar-specific H+ ATPase, a nucleic acid encoding vacuolar-specific H+ ATPase, or a compound that modulates vacuolar-specific H+ ATPase;
- and
- (c) cyclin-dependent kinase 2, a nucleic acid encoding cyclin-dependent kinase 2, or a compound that modulates cyclin-dependent kinase 2.
Dependencies are inferred from the claim text.