Surface transfection and expression procedure
The present invention relates to a method of transfecting cells comprising applying cells directly onto nucleic acids which are immobilized in transfection complexes on a surface and which transfect the cells. Preferably, the nucleic acids are immobilized in an array. In another aspect of the present invention, the method further includes expression of the nucleic acids in the transfected cells. In yet another aspect of the present invention, the method further comprises detecting the expression of the nucleic acids in the transfected cells.
1. A method, comprising:
a) providing:
i) a eukaryotic cell; and
ii) at least one transfection complex immobilized on a surface, said complex comprising first and second nucleic acids and first and second complexing agents, said first nucleic acid encoding a receptor protein and said second nucleic acid encoding a test protein, wherein said first and second nucleic acid are present in at least one expression vector, and said first complexing agent comprising a ligand for a cell surface receptor that is present on the surface of said eukaryotic cell, and said second complexing agent comprising a DNA binding molecule; and
b) contacting said eukaryotic cell with said complex under conditions such that the eukaryotic cell is co-transfected with said first and second nucleic acids and said first and second nucleic acids are expressed; and
c) detecting the presence or absence of binding between said receptor protein expressed by said first nucleic acid and said test protein expressed by said second nucleic acid.
2. The method of claim 1 , wherein said receptor protein is selected from the group consisting of G-protein coupled receptors and receptor kintses.
3. The method of claim 1 , wherein said at least one transfection complex immobilized on a surface form an array.
4. The method of claim 1 , wherein said detecting the presence or absence of binding between said receptor protein and said test protein comprises purifying said receptor protein and determining its mass.
5. The method of claim 1 , wherein the transfection complex further comprises a third complexing agent, said third complexing agent comprising a membrane permeable molecule.
6. The method of claim 5 , wherein the DNA-binding molecule is a cationic protein.
7. The method of claim 4 , wherein the member permeable molecule is a cationic lipid.
8. The method of claim 6 , wherein said ligand is covalently linked to the cationic protein.
9. The method of claim 6 , wherein said ligand is transferrin and the cationic protein is polylysine.
10. The method of claim 1 , wherein the transfection complex further comprises one or more additional complexing agents selected from the group consisting of targeting molecules, transcription molecules, nucleic acid degradation inhibitors, cell growth and integrity modulators, and mixtures thereof.