Method and system for incorporation of biomolecules into vesicles, cells, and micelles using electroactive porous device membrane

Patent metadata

Field	Value
Publication number	US-11261465-B2
Application number	US-201816026709-A
Country	US
Kind code	B2
Filing date	Jul 3, 2018
Priority date	Jul 5, 2017
Publication date	Mar 1, 2022
Grant date	Mar 1, 2022

Abstract

A method and system of delivering a charged cargo, such as a biomolecule, to a target structure, such as cells, exosomes, other vesicles or micelles, using an electroactive porous membrane. This method comprises contacting an electroactive porous membrane with a fluid flow toward the membrane. The fluid contains charged biomolecules and the membrane and biomolecules are oppositely charged so that the biomolecules in the fluid are trapped on the membrane as the fluid flows through the pores of the membrane. Acceptor cells of interest are pinned to the membrane by the flow of the fluid, thereby aggregating the cells onto the membrane in close proximity to the trapped biomolecules. Finally, the acceptor cells are permeabilized.

First claim

What is claimed is:

1. A method of controlled intracellular delivery of electrically charged biomolecules to target structures comprising:
embossing concavities into a surface of a device membrane;
providing electrically charged biomolecules to the device membrane;
positioning the electrically charged biomolecules at a surface of the device membrane by providing the device membrane with an electrical charge opposite to the electrically charged biomolecules;
directing the target structures to the surface of the device membrane by a fluid flow through the device membrane, which prevents passage of the target structures;
organizing the target structures on the device membrane using the concavities;
then releasing the electrically charged biomolecules from the surface of the device membrane by changing an electrical charge of the device membrane;
permeabilizing the target structures; and
delivering or allowing the electrically charged biomolecules to be delivered to the target structures.

2. The method of claim 1 , wherein the target structures are permeabilized by electroporation, sonoporation or by using the device membrane itself.

3. The method of claim 1 , wherein the electrically charged biomolecules are selected from the group consisting of plasmids, DNA and RNA and wherein the target structures are selected from the group consisting of cells, exosomes, liposomes and micelles.

4. The method of claim 1 , wherein the target structures are non-adherent cells.

5. The method of claim 1 , wherein the fluid flow pins the target structures to the device membrane thereby aggregating the target structures onto the device membrane in proximity to the charged biomolecules.

6. A genetic modification process comprising:
embossing concavities into a surface of a device membrane;
providing electrically charged biomolecules to the device membrane;
positioning electrically charged biomolecules at a surface of the device membrane by providing the device membrane with an electrical charge opposite to the electrically charged biomolecules;
directing target structures to the surface of the device membrane by a fluid flow and organizing the target structures on the device membrane using the concavities; and
then releasing the electrically charged biomolecules from the surface of the device membrane by changing an electrical charge of the device membrane and permeabilizing the target structures to deliver biomolecules to the target structures.

7. The process of claim 6 , wherein the target structures are permeabilized by electroporation, sonoporation or by using the device membrane itself.

8. The process of claim 6 , wherein the biomolecules are a charged material selected from the group consisting of plasmids, DNA and RNA and wherein the target structures are selected from the group consisting of cells, exosomes, liposomes and micelles.

9. The process of claim 6 , wherein the target structures are non-adherent cells.

Assignees

• Charles Stark Draper Laboratory Inc

Inventors

• Tandon Vishal
• Freeman Daniel K
• Coppeta Jonathan R
• Borenstein Jeffrey T
• Balestrini Jenna L

Classifications

• C12N15/87 Primary

  Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation · CPC title

• C12N13/00

  Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves · CPC title

• C12M35/02

  Electrical or electromagnetic means, e.g. for electroporation or for cell fusion · CPC title

• C12M1/42

  Apparatus for the treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves · CPC title