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US 5,633,156 AGranted
Methods for calcium phosphate transfection
AssigneeGenentech, Inc.
InventorsFlorian M. Wurm, Martin Jordan
Life of this patent
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Abstract
A method is provided for calcium phosphate transfection of a eukaryotic host cell wherein particles comprising calcium phosphate and a desired nucleic acid are grown to an optimal size and then contacted with the host cell under conditions providing a substantially slower particle growth rate, thereby increasing the host cell's exposure to optimally-sized particles.
Claims12
A method for introducing a desired nucleic acid into a eukeryotic host cell, comprising
- (a) admixing Ca.sup.2+, PO.sub.4.sup.3- and the desired nucleic acid to form a precipitation mixture;
- (b) incubating the precipitation mixture to form particles comprising calcium phosphate and the desired nucleic acid, and allowing the particles to grow to an average length of up to about 300 nm;
- (c) performing a step selected from the group consisting of:
- (1) diluting the precipitation mixture and simultaneously admixing the precipitation mixture with a eukaryotic host cell lacking a cell wall to form a transfection mixture;
- and (2) diluting the precipitation mixture to form a diluted precipitation mixture, and thereafter admixing the diluted precipitation mixture with a eukaryotic host cell lacking a cell wall to form a transfection mixture;
- and
- (d) incubating the transfection mixture to allow the eukaryoric host cell to take up the particles to form a transfected cell.
4 more elements
Dependencies are inferred from the claim text.