Method of inserting DNA into living cells

• US 4,394,448 A
• Filed: 04/24/1980
• Issued: 07/19/1983
• Est. Priority Date: 02/24/1978
• Status: Expired due to Term

• Current Assignee: Liposome Technology, Inc.
• Original Assignee: Liposome Technology, Inc.
• Inventors: Papahadjopoulos, Demetrios P., Szoka, Francis C. Jr.
• Primary Examiner(s): Marantz, Sidney
• Application Number:
  US06/143,455
• Time in Patent Office: 1,181 Days
• Field of Search: 435/172, 435/317, 424/19
• US Class Current: 435/458
• CPC Class Codes: A61K 9/127 Liposomes
• CPC Class Codes: C12N 15/88 using microencapsulation, e.g. using amphiphile liposome vesicle
• CPC Class Codes: Y10S 435/82 Subcellular parts of microorganisms

Abstract

A method of inserting deoxyribonucleic acid or fragments thereof into a living cell, which comprises; encapsulating the DNA or fragment in a lipid vesicle and bringing the vesicle in contact with said cell, whereby insertion occurs.

Claims

1. A method of inserting deoxyribonucleic acid into a living cell, which comprises;

encapsulating the acid material in a lipid vesicle and bringing the vesicle in contact with said cell, whereby insertion occurs.

4. The method of claim 1 wherein the living cell is an animal cell.

5. The method of claim 1 wherein the living cell is a unicellular microorganism.

6. The method of claim 5 wherein the microorganism is E. coli.

7. The method of claim 1 wherein the living cell is a plant cell.

8. The method of claim 1 wherein the living cell is a plant protoplast.

9. The method of claim 1 wherein the lipid vesicles are oligolamellar vesicles having an average diameter of 2,000 to 4,000 Angstroms.

2. A method of inserting deoxyribonucleic acid into a living cell, which comprises;

providing the deoxyribonucleic acid material encapsulated in a lipid vesicle; and

bringing the encapsulated material into contact with the cell, whereby insertion occurs.

3. A method of inserting deoxyribonucleic acid into a living cell, which comprises;

(1) providing a mixture of a vesicle wall forming compound in organic solvent and an aqueous mixture of the deoxyribonucleic acid material to be encapsulated, the ratio of organic phase to aqueous phase being that which will produce a water-in-oil emulsion;

(2) forming a homogeneous water-in-oil emulsion of said mixture, of the character produced by ultrasonic radiation;

(3) removing organic solvent from the emulsion, whereby a mixture is obtained having a gel-like character;

(4) converting the gel-like mixture to a suspension of synthetic, oligolamellar vesicles encapsulating the deoxyribonucleic acid material; and

bringing the encapsulated material into contact with the cell, whereby insertion occurs.

10. A method of inserting a chromosome into a living cell, which comprises;

providing the chromosome encapsulated in a lipid vesicle; and

bringing the lipid vesicle into contact with the cell, whereby insertion occurs.

11. A method of inserting a plasmid into a living cell, which comprises;

providing the plasmid encapsulated in a lipid vesicle; and

bringing the lipid vesicle into contact with the cell, whereby insertion occurs.