Transfection method and uses related thereto

• US 20030203486A1
• Filed: 03/28/2003
• Published: 10/30/2003
• Est. Priority Date: 09/17/1999
• Status: Abandoned Application

• Current Assignee: David M. Sabatini
• Original Assignee: David M. Sabatini
• Inventors: Sabatini, David M.
• Application Number:
  US10/403,720
• Publication Number:
  US 20030203486A1
• Time in Patent Office: Days
• US Class Current: 435/455
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
• CPC Class Codes: C12N 15/88 using microencapsulation, e.g. using amphiphile liposome vesicle
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2565/518 characterised by the immobilisation of the nucleic acid sample or target

Abstract

The invention features a method of introducing nucleic acid molecules into cells by (a) affixing a nucleic acid molecule-containing mixture onto the surface of a an object, wherein the nucleic acid molecule-containing mixture includes (i) nucleic acid molecules to be introduced into cells and (ii) a carrier protein; and (b) contacting cells and the affixed nucleic acid molecules under appropriate conditions for entry of the nucleic acid molecules into the cells, whereby the nucleic acid molecules are introduced into the cells.

Claims

1. A method of introducing nucleic acid molecules into cells, said method comprising the steps of:

(a) affixing a nucleic acid molecule-containing mixture onto the surface of an object selected from a sphere, fiber, rod, and tube, wherein said nucleic acid molecule-containing mixture comprises (i) nucleic acid molecules to be introduced into cells and (ii) a carrier; and

(b) contacting cells and said affixed nucleic acid molecules under appropriate conditions for entry of said nucleic acid molecules into said cells, whereby said nucleic acid molecules are introduced into said cells.

2. The method of claim 1, wherein said nucleic acid molecule-containing mixture further comprises an appropriate transfection reagent.

3. The method of claim 1, wherein said nucleic acid molecule-containing mixture further comprises a buffer that facilitates nucleic acid molecule condensation.

4. The method of claim 1, wherein each nucleic acid molecule to be introduced is contained in a vector.

5. The method of claim 4, wherein said vector is a plasmid or a viral-based vector.

6. The method of claim 1, wherein said nucleic acid molecules are expressed in said cells.

7. The method of claim 1, wherein said nucleic acid molecules comprise double-stranded RNA molecules or DNA molecules encoding double-stranded RNA molecules.

8. The method of claim 1, wherein said cells are mammalian cells.

9. The method of claim 1, wherein said nucleic acid molecules are DNA or RNA molecules.

10. The method of claim 1, wherein said carrier is a gelatin.

11. The method of claim 10, wherein said gelatin is selected from the group consisting of a protein gelatin, a hydrogel, a sugar-based gelatin, and a synthetic gelatin.

12. A composition comprising (i) an object selected from a sphere, fiber, rod, and tube, (ii) nucleic acid molecules to be introduced into cells, and (ii) a carrier;

wherein said nucleic acid molecules and said carrier are affixed to said object such that, under appropriate conditions for entry of said nucleic acid molecules into cells, said nucleic acid molecules but not said object enter into said cells.

13. The composition of claim 12, wherein said composition further comprises an appropriate transfection reagent.

14. The composition of claim 12, wherein said nucleic acid molecule-containing mixture further comprises a buffer that facilitates nucleic acid molecule condensation.

15. The composition of claim 12, wherein each nucleic acid molecule to be introduced is contained in a vector.

16. The composition of claim 15, wherein said vector is a plasmid or a viral-based vector.

17. The composition of claim 12, wherein said nucleic acid molecules comprise double-stranded RNA molecules or DNA molecules encoding double-stranded RNA molecules.

18. The composition of claim 12, wherein said nucleic acid molecules are DNA or RNA molecules.

19. The composition of claim 12, wherein said carrier is a gelatin.

20. The composition of claim 19, wherein said gelatin is selected from the group consisting of a protein gelatin, a hydrogel, a sugar-based gelatin, and a synthetic gelatin.