Method for selecting improved vectors
OwnerAlan John Kingsman, Jason Slingsby +1inventors
InventorsAlan John Kingsman, Jason Slingsby, Melvyn Yap
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Abstract
A method is provided for selecting an improved retroviral genome having an improved packaging efficiency which method comprises: a) introducing one or more random mutations into a retroviral genome comprising a packaging signal; b) introducing the mutagenised retroviral genome into a host cell expressing viral polypeptides required for packaging the retroviral genome; c) determining whether retroviral packaging efficiency in the cell is improved as compared with a retroviral genome comprising a non-mutated packaging signal; d) selecting a viral genome which has improved packaging efficiency.
Claims21
A method for selecting an improved retroviral genome having an improved packaging efficiency which method comprises:
- (a) introducing one or more random mutations into a retroviral genome comprising a packaging signal;
- (b) introducing the mutagenised retroviral genome into a host cell expressing viral polypeptides required for packaging the retroviral genome;
- (c) determining whether retroviral packaging efficiency in the cell is improved as compared with a retroviral genome comprising a non-mutated packaging signal;
- (d) selecting a viral genome which has improved packaging efficiency.
A method for enhancing the efficiency of retroviral packaging which method comprises expressing in a producer cell at least a first nucleotide sequence encoding a retroviral gag-pol polypeptide, a second nucleotide sequence encoding a retroviral envelope polypeptide and a third nucleotide sequence encoding a retroviral genome wherein the ratio of the first nucleotide sequence to the second nucleotide sequence and the ratio of the first nucleotide sequence to the third nucleotide sequence is x:y and x:z, respectively wherein x is less than 1 and y and z are 1.
A producer cell which expresses at least a first nucleotide sequence encoding a retroviral gag-pol polypeptide, a second nucleotide sequence encoding a retroviral envelope polypeptide and a third nucleotide sequence encoding a retroviral genome wherein the ratio of the first nucleotide sequence to the second nucleotide sequence and the ratio of the first nucleotide sequence to the third nucleotide sequence is x:y and x:z, respectively wherein x is less than 1 and y and z are 1.
Dependencies are inferred from the claim text.