Vectors having enhanced expression, and methods of making and uses thereof
First Claim
1. A vector for enhanced expression of at least one first nucleic acid molecule in a cell, said vector modified to comprise nucleic acid sequences comprising the first nucleic acid molecule and at least one second nucleic acid molecule encoding a translation factor, wherein the vector is modified to comprise the nucleic acid sequences by at least one insertion into the vector, and there is substantially co-temporal expression of the first and second nucleic acid molecules, whereby expression of the second nucleic acid molecule enhances expression of the first nucleic acid molecule by enhancing translation, wherein the insertion is not present in the vector prior to the modification.
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Accused Products
Abstract
Disclosed and claimed are vectors having enhanced expression and methods for making and using them. Enhancement of expression is from substantially co-temporal expression of at least one first nucleic acid molecule and at least one second nucleic acid molecule. The second nucleic acid molecule encodes a translation factor. The contemporaneous expression can be from operably linking the first and second nucleic molecules to a single promoter, or from operably linking the first nucleic acid molecule to a first promoter and the second nucleic molecule to a second promoter wherein the first and second promoters function substantially contemporaneously. Thus, the first and second nucleic acid molecules can be at the same locus in the vector or at different loci. The second nucleic acid molecule can encode encode one translation factor or more than one translation factor. The translation factor can be a K3L open reading frame, an E3L open reading frame, a VAI RNA, an EBER RNA, a sigma 3 open reading frame, a TRBP open reading frame, or combinations thereof. The vector can be a poxvirus such as an attenuated poxvirus; for instance, a NYVAC vaccinia virus or an ALVAC canarypox virus.
100 Citations
15 Claims
- 1. A vector for enhanced expression of at least one first nucleic acid molecule in a cell, said vector modified to comprise nucleic acid sequences comprising the first nucleic acid molecule and at least one second nucleic acid molecule encoding a translation factor, wherein the vector is modified to comprise the nucleic acid sequences by at least one insertion into the vector, and there is substantially co-temporal expression of the first and second nucleic acid molecules, whereby expression of the second nucleic acid molecule enhances expression of the first nucleic acid molecule by enhancing translation, wherein the insertion is not present in the vector prior to the modification.
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15. A method of increasing expression of at least one first nucleic acid molecule by a vector comprising the first nucleic acid molecule, in a cell, comprising modifying the vector to comprise at least one second nucleic acid molecule encoding a translation factor, and expressing the vector in the cell;
- wherein there is substantially co-temporal expression of the first and second nucleic acid molecules, whereby expression of the second nucleic acid molecule enhances expression of the first nucleic acid molecule by enhancing translation.
Specification