Device and method for electroporation-based delivery of molecules into cells and dynamic monitoring of cell responses
The present invention includes devices and methods for transfecting a cell or cell population and dynamic monitoring of cellular events. A variety of microelectronic devices are provide that incorporate functions such as electroporation, modulation of a transmembrane potential and dynamic monitoring of cellular functions and mechanisms.
1. A method of monitoring a cellular response in real time, comprising:
a. monitoring at least one impedance measurement prior to transfecting a cell or cell population;
b. determining said cell or cell population is to be transfected if said at least one impedance measurement prior to transfecting said cell or cell population is within a predetermined range;
c. transfecting said cell or cell population with a molecule if determined to be transfected, wherein said cell or cell population is transfected using a method selected from the group consisting of a chemical transfection method, electroporation and a viral transfection method, further wherein said molecule is capable of affecting a cellular function selected from the group consisting of cell proliferation, cell adhesion and cell spreading; and
d. monitoring impedance of said cell or said cell population when said cell or said cell population is attached to an impedance-monitoring electrode.
2. The method according to claim 1 , wherein said cell is a eukaryotic cell.
3. The method according to claim 1 , wherein said cell is a mammalian cell.
4. The method according to claim 1 , wherein said molecule is selected from the group consisting of a DNA molecule, an RNA molecule, an oligopeptide, a polypeptide, a protein and a compound.
5. The method according to claim 4 , wherein said DNA molecule is selected from the group consisting of a recombinant DNA molecule, a native DNA molecule, a plasmid, a cDNA, an anti-sense DNA strand and an oligonucleotide.
6. The method according to claim 4 , wherein said RNA molecule is selected from the group consisting of a siRNA molecule, a microRNA molecule, a native RNA molecule, a ribozyme RNA and an aptamer.
7. The method according to claim 1 , wherein said molecule is a capable of affecting a process selected from the group consisting of transcription, translation, RNA splicing and RNA editing.
8. The method according to claim 1 , wherein said molecule is capable of affecting cell morphology.
9. The method according to claim 1 , further comprising conducting an endpoint assay selected from the group consisting of a cell viability assay, an apoptosis assay, an enzymatic activity assay, a signal transduction analysis assay, and a reporter assay.
10. A method of monitoring a cellular response in real time, comprising:
a. monitoring at least one impedance measurement prior to transfecting a cell or cell population;
b. determining said cell or cell population is to be transfected if said at least one impedance measurement prior to transfecting said cell or cell population is within a predetermined range;
c. transfecting said cell or cell population with a molecule if determined to be transfected, wherein said cell or cell population is transfected using a method selected from the group consisting of a chemical transfection method, electroporation and a viral transfection method, further wherein said molecule is capable of affecting a cellular receptor or a signal transduction pathway activated by said receptor; and
d. monitoring impedance of said cell or said cell population when said cell or said cell population is attached to an impedance-monitoring electrode.
11. The method according to claim 10 , further comprising conducting an endpoint assay selected from the group consisting of a cell viability assay, an apoptosis assay, an enzymatic activity assay, a signal transduction analysis assay, and a reporter assay.
12. The method according to claim 10 , wherein said cell is a eukaryotic cell.
13. The method according to claim 10 , wherein said cell is a mammalian cell.
14. The method according to claim 10 , wherein said molecule is selected from the group consisting of a DNA molecule, an RNA molecule, an oligopeptide, a polypeptide, a protein and a compound.
15. The method according to claim 14 , wherein said DNA molecule is selected from the group consisting of a recombinant DNA molecule, a native DNA molecule, a plasmid, a cDNA, an anti-sense DNA strand and an oligonucleotide.
16. The method according to claim 14 , wherein said RNA molecule is selected from the group consisting of a siRNA molecule, a microRNA molecule, a native RNA molecule, a ribozyme RNA and an aptamer.
17. The method according to claim 10 , wherein said molecule is a capable of affecting a process selected from the group consisting of transcription, translation, RNA splicing and RNA editing.
18. The method according to claim 10 , wherein said molecule is capable of affecting cell morphology.
19. A method of monitoring a cellular response in real time, comprising:
a. transfecting a cell or cell population with a molecule using electroporation, wherein said molecule is capable of affecting a cellular function selected from the group consisting of cell proliferation, cell adhesion, and cell spreading; and
b. monitoring impedance of said cell or said cell population when said cell or said cell population is attached to an impedance-monitoring electrode.
20. The method according to claim 19 , wherein said cell is a mammalian cell.
21. The method according to claim 19 , wherein said molecule is selected from the group consisting of a DNA molecule, a RNA molecule and a polypeptide.
22. The method according to claim 21 , wherein said DNA molecule is selected from the group consisting of a recombinant DNA molecule, a native DNA molecule, a plasmid, a cDNA, an anti-sense DNA strand and an oligonucleotide; and wherein said RNA molecule is selected from the group consisting of a siRNA molecule, a microRNA molecule, a native RNA molecule, a ribozyme RNA and an aptamer.
23. A method of monitoring a cellular response in real time, comprising:
a. transfecting a cell or cell population with a molecule using electroporation, wherein said molecule is capable of affecting cell morphology; and
b. monitoring impedance of said cell or said cell population when said cell or said cell population is attached to an impedance-monitoring electrode.
24. The method according to claim 23 , wherein said cell is a mammalian cell.
25. The method according to claim 23 , wherein said molecule is selected from the group consisting of a DNA molecule, a RNA molecule and a polypeptide.
26. The method according to claim 25 , wherein said DNA molecule is selected from the group consisting of a recombinant DNA molecule, a native DNA molecule, a plasmid, a cDNA, an anti-sense DNA strand and an oligonucleotide; and wherein said RNA molecule is selected from the group consisting of a siRNA molecule, a microRNA molecule, a native RNA molecule, a ribozyme RNA and an aptamer.
27. A method of monitoring a cellular response in real time, comprising:
a. transfecting a cell or cell population with a molecule using electroporation, wherein said molecule is capable of affecting a cellular receptor or a signal transduction pathway activated by said receptor; and
b. monitoring impedance of said cell or said cell population when said cell or said cell population is attached to an impedance-monitoring electrode.
28. The method according to claim 27 , wherein said cell is a mammalian cell.
29. The method according to claim 27 , wherein said molecule is selected from the group consisting of a DNA molecule, a RNA molecule and a polypeptide.
30. The method according to claim 29 , wherein said DNA molecule is selected from the group consisting of a recombinant DNA molecule, a native DNA molecule, a plasmid, a cDNA, an anti-sense DNA strand and an oligonucleotide; and wherein said RNA molecule is selected from the group consisting of a siRNA molecule, a microRNA molecule, a native RNA molecule, a ribozyme RNA and an aptamer.