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US 2003/0166279 A1Published application
Minicell-based transfection
OwnerRoger A. Sabbadini, Neil Berkleyinventors
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Abstract
The invention provides compositions and methods for the production of achromosomal and anucleate cells useful for applications such as diagnositic and therapeutic uses, as well as research tools and agents for drug discovery.
Claims18
A method of determining the rate of transfer of nucleic acid from a minicell to a cell, comprising
- (a) contacting said cell to said minicell, wherein said minicell comprises said nucleic acid, for a measured period of time;
- (b) separating minicells from said cells;
- (c) measuring the amount of nucleic acid in said cells, wherein the amount of nucleic acid in said cells over said measured period of time is the rate of transfer of a nucleic acid from a minicell.
A method of determining the amount of a nucleic acid transferred to a cell from a minicell, comprising
- (a) contacting said cell to said minicell, wherein said minicell comprises an expression element having eukaryotic expression sequences operably linked to an ORF encoding a detectable polypeptide, wherein said minicell displays a binding moiety, and wherein said binding moiety binds an epitope of said cell;
- and
- (b) detecting a signal from said detectable polypeptide, wherein a change in said signal corresponds to an increase in the amount of a nucleic acid transferred to a cell.
A method of detecting the expression of an expression element in a cell, comprising
- (a) contacting said cell to a minicell, wherein said minicell comprises an expression element having cellular expression sequences operably linked to an ORF encoding a detectable polypeptide, wherein said minicell displays a binding moiety, and wherein said binding moiety binds an epitope of said cell;
- (b) incubating said cell and said minicell for a period of time effective for transfer of nucleic acid from said minicell to said cell;
- and
- (c) detecting a signal from said detectable polypeptide, wherein an increase in said signal corresponds to an increase in the expression of said expression element.
1 more element
Dependencies are inferred from the claim text.