Oligonucleotide-facilitated coalescence

• US 20040013654A1
• Filed: 08/12/2003
• Published: 01/22/2004
• Est. Priority Date: 10/12/2000
• Status: Active Grant

• Current Assignee: Rutgers University
• Original Assignee: University of Medicine and Dentistry of New Jersey (State of New Jersey)
• Inventors: Kramer, Fred R, Alsmadi, Osama A, Tyagi, Sanjay
• Granted Patent: US 7,129,087 B2
• Time in Patent Office: Days
• US Class Current: 424/93.21
• CPC Class Codes: A61K 9/127 Liposomes
• CPC Class Codes: A61K 9/1271 Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers liposome as conjugate A61K47/6911
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Abstract

Coalescence of cells or other membrane-bound entities is facilitated by anchoring an outwardly projecting first oligonucleotide in one member and an outwardly projecting second oligonucleotide, complementary to the first, in a second member and incubating under hybridizing conditions. Liposomes may be coalesced with cells to deliver hydrophilic agents thereto, such as DNA probes or drugs. Kits containing complementary oligonucleotides containing hydrophobic anchoring moieties may be used.

Claims

1. A method for facilitating the coalescence of two members selected from the group consisting of cells, vesicles, liposomes and lipidic particles comprising:

a. incorporating in the first of said members a first outwardly projecting anchored oligonucleotide containing a first terminal hydrophobic anchoring moiety,

b. incorporating in the second of said members a second outwardly projecting anchored oligonucleotide complementary to said first anchored oligonucleotide and also containing a second terminal hydrophobic anchoring moiety, and

c. incubating said first and second members under conditions promoting hybridization of said first and second outwardly projecting anchored oligonucleotides.

2. The method according to claim 1, wherein said first and second members are cells.

3. The method according to claim 1, wherein the first member is a cell and the second member is a liposome or lipidic particle containing an entrapped hydrophilic substance to be inserted into said cell.

4. The method according to claim 3, wherein said hydrophilic substance is selected from the group consisting of DNA, drugs, therapeutic agents, chemotherapeutic agents, antisense agents, stains and oligonucleotide probes.

5. The method according to claim 1, wherein each of the first and second and second terminal hydrophobic anchoring moieties is selected from the group consisting of cholesterol, a fatty acid, a hydrophobic peptide and a lipid.

6. The method according to claim 1, wherein at least one of said first and second terminal hydrophobic anchoring moieties is a hemifusion destabilizer.

7. The method according to claim 1, wherein said incubation is carried out in the presence of a hemifusion destabilizing agent.

8. A liposome or lipidic particle containing an entrapped hydrophilic substance and having incorporated therein an outwardly projecting anchored oligonucleotide containing a terminal hydrophobic anchoring moiety.

9. The liposome or lipidic particle of claim 8, wherein the anchoring moiety is a hemifusion destabilizer.

10. The liposome or lipidic particle of claim 8, wherein the entrapped hydrophilic substance is selected from the group consisting of DNA, drugs, therapeutic agents, chemotherapeutic agents, antisense agents, stains and oligonucleotide probes.

11. A kit of reagents comprising a pair of separately packaged complementary oligonucleotides each containing a terminal hydrophobic anchoring moiety.

12. The kit according to claim 11 further comprising a hemifusion destabilizing agent.