Method and apparatus for avalanche-mediated transfer of agents into cells
View Patent ↗The present invention provides a method and apparatus for transferring an agent into a cell. The method includes the steps of providing an agent outside of a cell and generating a vapor bubble and a plasma discharge between an avalanche electrode and a conductive fluid surrounding the cell. The vapor bubble and plasma discharge generate a mechanical stress wave and an electric field, respectively. The combination of this mechanical stress wave and electric field results in permeabilization of the cell, which in turn results in transfer of the agent into the cell.
1. A method of directly transferring a polypeptide or an RNA molecule into a cell, comprising:
(a) providing a polypeptide or an RNA molecule outside a cell; and
(b) generating an electric field and a mechanical stress wave by applying a voltage to an electrode to form a plasma discharge, the electrode providing both the electric field and the mechanical stress wave to the cell, thereby directly transferring the polypeptide or the RNA molecule into the cell.
2. The method of claim 1 , wherein the electric field and mechanical stress wave are generated by applying voltage pulses at a frequency of between about 0.1 Hz and about 1 kHz.
3. The method of claim 1 , wherein generating the electric field and the mechanical stress wave comprises generating a non-uniform electric field between the electrode and a return electrode, wherein the electric field around the electrode is sufficient for generating a plasma discharge.
4. The method of claim 1 , wherein the cell is a eukaryotic cell, a prokaryotic cell, a primary cell, a cell line, or is part of a tissue.
5. The method of claim 1 , wherein generating the electric field and the mechanical stress wave permeabilizes the cell.
6. The method of claim 1 , wherein generating the electric field and the mechanical stress wave comprises generating the electric field and the mechanical stress wave substantially concurrently.
7. The method of claim 1 , wherein the plasma discharge formed is a transient plasma discharge near the cell.
8. The method of claim 2 , wherein the cell is a eukaryotic cell, a prokaryotic cell, a primary cell, a cell line, or is part of a tissue.
9. The method of claim 2 , wherein generating the electric field and the mechanical stress wave permeabilizes the cell.
10. The method of claim 7 , wherein the cell is a eukaryotic cell, a prokaryotic cell, a primary cell, a cell line, or is part of a tissue.
11. The method of claim 7 , wherein forming the transient plasma discharge permeabilizes the cell.
12. A method of transferring a polypeptide or an RNA molecule into a cell, comprising:
(a) providing a polypeptide or an RNA molecule outside a cell; and
(b) generating an electric field and a mechanical stress wave by applying a voltage in a range of about 100 V to about 10 kV to an electrode to form a plasma discharge, the electrode providing both the electric field and the mechanical stress wave to the cell, thereby transferring the polypeptide or the RNA molecule into the cell.
13. The method of claim 1 , wherein the electrode is positioned at a distance of about 0.01 mm to about 1 cm from the cell.
14. The method of claim 2 , wherein the voltage is applied in a range of about 100 V to about 10 kV.
15. The method of claim 2 , wherein the electrode is positioned at a distance of about 0.01 mm to about 1 cm from the cell.
16. The method of claim 12 , wherein the plasma discharge formed is a transient plasma discharge near the cell.
17. A method of transferring a polypeptide or an RNA molecule into a cell, comprising:
(a) providing the polypeptide or the RNA molecule outside of the cell; and
(b) transferring the polypeptide or the RNA molecule into the cell by applying voltage to an electrode to form a transient plasma discharge near the cell, wherein the electrode is positioned at a distance of about 0.01 mm to about 1 cm from the cell, the electrode providing both an electric field and a mechanical stress wave to the cell.