Method for intracellular modifications within living cells using pulsed electric fields

A method of regulating transcription of a gene in a eukaryotic cell, comprising:

• selecting at least one gene selected from the group consisting of ITPKA, AHNAK, EMP3, ADORA2B, POU2AF1, AIM1, ATP1G1, ASNS, ETS2, CD45,
• VIM, TGIF, LAT, CLIC4, SLC7A5, ZFP36L2, RUNX1, SLC3A2, IFRD1, and PrP;
• and
• applying at least one electric field pulse to the cell, wherein each electric field pulse has a pulse duration of less than about 100 nanoseconds.

A method of determining induction of gene transcription in response to electropertubation, comprising:

• suspending a plurality of cells in a medium;
• applying at least one electric field pulse to said plurality of cells, wherein each electric field pulse has a pulse duration of less than about 100 nanoseconds;
• identifying at least one cell which is electroperturbed;
• isolating said electroperturbed cell;
• and
• determining cellular gene transcription in said electroperturbed cell.

A method of marking a eukaryotic cell for diagnostic or therapeutic procedures, comprising:

• suspending a plurality of cells in a medium;
• electroperturbing said plurality of cells, thereby inducing a cellular response in at least a portion of the cells, wherein said cellular response marks said at least a portion of the cells for a diagnostic or therapeutic procedure;
• and
• identifying said at least a portion of the cells that were electroperturbed by the presence of said cellular response.

A method of determining cellular tolerance to electropertubation, comprising:

• (a) suspending one or more cells in a medium;
• (b) applying a first electric field pulse to one or more cells,
• (c) identifying electroperturbed cells;
• (d) isolating said electroperturbed cells;
• (e) identifying one or more indicators of cellular response in said electroperturbed cells;
• (f) applying a second electric field pulse to one or more cells;
• (g) repeating steps (c)-(e)
• (h) comparing said one or more indicators of cellular response after application of the first electric field with said one or more indicators of cellular response after application of the second electric field.

A method of selectively electroperturbing a population of cells, comprising:

• determining a dielectric property of one or more cells in a first sub-population of cells;
• determining a dielectric property of one or more cells in a second population of cells;
• determining an electric field pulse based on said dielectric property of said first sub-population of cells and said dielectric property of said second population of cells, wherein said electric field pulse selectively electroperturbs the first sub-population of cells without substantially affecting the second population of cells.
• obtaining a cell suspension, wherein said cell suspension contains said first sub-population of cells and said second population of cells;
• and
• applying said electric field pulse to said cell suspension, thereby electroperturbing said first sub-population of cells without substantially affecting the second population of cells.

A method of reducing proliferation of rapidly dividing cells in a patient, comprising;

• removing a portion of a patient's tissue, wherein said tissue contains rapidly dividing cells and terminally differentiated cells;
• applying at least one electric field pulse to one or more cells in said tissue, wherein each electric field pulse has a pulse duration of less than about 100 nanoseconds;
• and
• reintroducing said tissue into said patient.

A method of treating a tumor in a patient, comprising;

• identifying a tumor in the patient;
• applying a catheterized electrode to said patient proximate to said tumor;
• wherein said catheterized electrode is capable of providing at least one electric field pulse;
• and
• applying said at least one electric field pulse to at least a portion of said tumor, wherein each electric field pulse has a pulse duration of less than about 100 nanoseconds, thereby treating said tumor.

A method of facilitating entry of a diagnostic or therapeutic agent into a cell's intracellular structures, comprising:

• applying at least one first electric field pulse to the cell, said first electric pulse sufficient to cause electroporation;
• incubating said cell with the therapeutic agent;
• and
• applying one or more second electric field pulses to one or more cells in said tissue, wherein each second electric field pulse has a pulse duration of less than about 100 nanoseconds.