Method and articles for transfection of genetic material

• US 6,475,994 B2
• Filed: 12/23/1998
• Issued: 11/05/2002
• Est. Priority Date: 01/07/1998
• Status: Expired due to Fees

• Current Assignee: Dendritic NanoTechnologies, Inc. (Loveland Products, Inc.)
• Original Assignee: Donald A. Tomalia
• Inventors: Balogh, Lajos, Tomalia, Donald A.
• Primary Examiner(s): NGUYEN, DAVE TRONG
• Application Number:
  US09/219,907
• Publication Number:
  US 20020013283A1
• Time in Patent Office: 1,413 Days
• Field of Search: 435/320.1, 435/69.1, 435/325, 435/455, 435/458, 514/44, 424/486, 424/497
• US Class Current: 514/44.00R
• CPC Class Codes: B01J 13/02 Making microcapsules or microballoons for medical preparations A61K9/50
• CPC Class Codes: C08G 83/003 Dendrimers
• CPC Class Codes: C12N 15/895 using biolistic methods
• CPC Class Codes: Y10S 977/914 Protein engineering
• CPC Class Codes: Y10S 977/916 Gene therapy

Abstract

A gene transfection particle includes a polymer, a support particle conjugated with the dendritic polymer, and genetic material conjugated with the dendritic polymer. The gene transfection particles are highly efficient and are capable of delivering higher quantities of genetic materials to cells, with reduced cell damage. A gene transfection method involves bombarding cells with conjugates of polymers and genetic material, with or without a support particle.

Claims

1. A gene transfection particle comprising a composite material including:

a dendritic polymer; and

a metal selected from the group consisting of one or more gold, silver, copper, magnesium, or calcium particles, gold sols, gold atoms, gold ions, gold containing complexes or molecules, and clusters or a mixture thereof conjugated with the dendritic polymer, wherein the maximum dimension of the conjugate is from about 1 nm to about 1000 nm; and

the conjugate further conjugated to genetic material.

2. The particle of claim 1, wherein the dendritic polymer comprises amine functional groups.

3. The particle of claim 2, wherein the dendritic polymer is a PAMAM, POPAM or PEI.

4. The particle of claim 2, wherein the dendritic polymer is a PAMAM.

5. The particle of claim 1, wherein the genetic material is DNA, a DNA fragment, or an oligonucleotide.

6. The particle of claim 5, wherein the dendritic polymer comprises amine functional groups.

7. The particle of claim 6, wherein the dendritic polymer is a PAMAM.

8. The particle of claim 1, wherein the metal is in the form of a single metal atom or atoms or ions, or compounds containing a single metal atom.

9. The particle of claim 8, wherein the dendritic polymer is a polyamine.

10. The particle of claim 8, wherein the dendritic polymer is a PAMAM, POPAM or PEI.

11. The particle of claim 8, wherein the dendritic polymer is a PAMAM.

12. A gene transfection particle comprising:

a metal selected from the group consisting of one or more gold, silver, copper, magnesium, or calcium particles, gold sols, gold atoms, gold ions, gold containing complexes or molecules, and clusters or a mixture thereof;

and a dendritic polymer attached to the surface of the metal, wherein the maximum dimension of the metal and dendritic polymer conjugate is from about 1 nm to about 1000 nm; and

genetic material conjugated to the metal and dendritic polymer conjugate.

13. The gene transfection particle of claim 12, wherein the maximum dimension of the gene transfection particle is from about 1 nm to about 1000 nm.

14. The gene transfection particle of claim 12, wherein the maximum dimension of the gene transfection particle is from about 1 nm to about 100 nm.

15. A method for delivering genetic material to plant or animal cells comprising:

conjugating a dendritic polymer to a metal selected from the group consisting of one or more gold, silver, copper, magnesium, or calcium particles, gold sols, gold atoms, gold ions, gold containing complexes or molecules, and clusters or a mixture thereof to form a polymer-metal conjugate, wherein the maximum dimension of the conjugate is from about 1 nm to about 1000 nm;

conjugating genetic material to the polymer-metal conjugate to form a gene transfection particle; and

accelerating the gene transfection particle toward a plant or animal cell with sufficient motive force to cause the gene transfection particle to penetrate and enter the cell.

16. The method of claim 15, wherein the gene transfection particle is accelerated with a high-voltage electric discharge.

17. The method of claim 15, wherein the gene transfection particle is accelerated with pressure discharge.

18. The method of claim 15, wherein the gene transfection particle is accelerated into a cell in vivo.