Back to results
US 8,772,008 B2Granted
Methods and compositions for increasing nuclease activity
AssigneeSangamo BioSciences, Inc.
InventorYannick Doyon
Life of this patent
Estimating the term…
Abstract
Methods and compositions for increasing nuclease activity by subjecting cells expressing the nuclease to hypothermic conditions to increase activity of the nucleases for genomic modifications.
Claims4
A method for increasing the nuclease activity of at least one zinc finger nuclease in an isolated mammalian cell, the method comprising:
- (a) culturing the isolated mammalian cell at an optimal growth temperature above 33° C.;
- (b) transfecting the isolated mammalian cell with a polynucleotide encoding the at least one zinc finger nuclease (ZFN) into the isolated mammalian cell from step (a);
- (c) culturing the isolated mammalian cell from step (b) at a temperature between 27° C. and 33° C. for between 1 and 4 days;
- and
- (d) culturing the isolated mammalian cell following step (c) at the optimal growth temperature, such that the nuclease activity of the ZFN is increased as compared to an isolated mammalian cell transfected and cultured only at the optimal growth temperature without the culturing step (c).
2 more elements
Dependencies are inferred from the claim text.