Methods of packaging multiple adeno-associated virus vectors
AssigneeUniversity of Florida Research Foundation, Inc.
InventorsBarry John Byrne, Phillip A. Doerfler, Nathalie Clement
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Abstract
Provided herein are methods related to co-packaging of multiple rAAV particles, e.g., by introducing multiple nucleic acid vectors encoding proteins or polypeptides or RNAs of interest into a single cell preparation.
Claims21
A method of producing a recombinant adeno-associated virus (rAAV) particle preparation having a desired ratio of a first rAAV particle to a second rAAV particle, the method comprising:
- contacting a producer cell preparation with:
- (i) a first nucleic acid vector containing a first construct comprising a heterologous nucleic acid region encoding a first protein or polypeptide, and
- (ii) a second nucleic acid vector containing a second construct comprising a heterologous nucleic acid region encoding a second protein or polypeptide;
- and
- culturing the producer cell preparation with the first and second nucleic acid vectors for a period of time sufficient to result in the production of a heterogeneous mixture of rAAV particles comprising a first rAAV particle comprising the first construct and a second rAAV particle comprising the second construct,
- wherein the heterogeneous mixture of rAAV particles has a ratio of the first rAAV particle to the second rAAV particle that is approximately the same as an input ratio of the first nucleic acid vector to the second nucleic acid vector.
A method of producing a recombinant adeno-associated virus (rAAV) particle preparation having a desired ratio of a first rAAV particle to a second rAAV particle, the method comprising:
- transfecting a producer cell preparation with at least a first plasmid encoding a first protein or polypeptide and a second plasmid encoding a second protein or polypeptide;
- and
- culturing the producer cell preparation with the first and second plasmids for a period of time sufficient to result in the production of a heterogeneous mixture of rAAV particles comprising a first rAAV particle comprising the first protein or polypeptide and a second rAAV particle comprising the second protein or polypeptide,
- wherein the heterogeneous mixture of rAAV particles has a ratio of the first rAAV particle to the second rAAV particle that is approximately the same as an input ratio of the first plasmid to the second plasmid.
A method of producing a recombinant adeno-associated virus (rAAV) particle preparation having a desired ratio of a first rAAV particle to a second rAAV particle, the method comprising:
- infecting a producer cell preparation with at least a first viral particle comprising a first nucleic acid vector encoding a first protein or polypeptide and a second viral particle comprising a second nucleic acid vector encoding a second protein or polypeptide;
- and
- culturing the producer cell preparation with the first and second viral particles for a period of time sufficient to result in the production of a heterogeneous mixture of rAAV particles comprising a first rAAV particle comprising the first protein or polypeptide and a second rAAV particle comprising the second protein or polypeptide,
- wherein the heterogeneous mixture of rAAV particles has a ratio of the first rAAV particle to the second rAAV particle that is approximately the same as an input ratio of the first nucleic acid vector to the second nucleic acid vector.
Dependencies are inferred from the claim text.