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US5766901A
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Apparatus and method for delivering a nucleotide into cell nuclei
Assignee: Leland Stanford Junior University
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Filing: 1995-05-04
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Granted: 1998-06-16
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Expires: 2015-05-04
Abstract
Abstract The invention presents a system for inducing cells in living intact tissue, in vivo or ex vivo, to accept nucleotides from their extracellular environment and to localize those nucleotides into the cells' nuclei. This system relies on the fact that, when subjected to high pressure, cells take in nucleotides and localize those nucleotides into their nuclei with a transfection rate of greater than 90% in some cases. This invention employs various techniques for placing under high pressure either cells in isolated tissue cultures, or cells in tissues still connected to a living body. Such pressurization techniques include enclosing the tissue in an impermeable and inelastic sheath and injecting fluid into the tissue until a sufficiently high pressure is reached; placing a tissue culture in a dish and placing the dish in a pressurized chamber; mechanical pressurization techniques such as compression of a space containing fluids; occluding the ingress and egress of a vessel or organ to create a watertight compartment within the vessel or organ; and pressurization of an entire organism after delivery of nucleotide to the extracellular environment. This invention also employs various techniques for delivering a solution or suspension containing the nucleotide into the tissue; such techniques include the use of a syringe-like mechanism and a catheter.
Inventors
- 👤 Michael J. Mann
- 👤 Frank P. Diet
- 👤 Victor J. Dzau
- 👤 Gary H. Gibbons
- 👤 Heiko Von der Leyen
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 16:41
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/11; C12N15/111; A; A61; A61M; A61M37/00; A61M5/00; A61M5/14; A61M5/142; A61M5/145; C12M; C12M23/00; C12M23/02; C12M23/14; C12M35/00; C12M35/04; C12M41/00; C12M41/40; C12N15/113; C12N15/1136; C12N15/87; C12N15/89; A61M25/00; A61M25/10; A61M2025/1043; A61M2025/105; A61M2025/1052; A61M25/1011; C12N2310/00; C12N2310/30; C12N2310/35; C12N2310/351; C12N2310/3517; C12N2320/00; C12N2320/30; C12N2320/32
Claims Analysis
Total Claims: 25
Claim 1
Independent Claim
61 words
A method of delivering a nucleic acid molecule into a cell, said method comprising the steps of: contacting said cell with a nucleic acid molecule in a liquid medium; establishing a sealed enclosure around said cell and said liquid medium; and establishing in said sealed enclosure an incubation pressure sufficient to enhance uptake of said nucleic acid molecule by said cell.
Claim 2
Independent Claim
61 words
A method of delivering a nucleic acid molecule into a cell, said method comprising the steps of: contacting said cell with a nucleic acid molecule in a liquid medium; establishing a sealed enclosure around said cell and said liquid medium; and establishing in said sealed enclosure an incubation pressure sufficient to enhance uptake of said nucleic acid molecule by said cell.
Claim 3
Dependent Claim
14 words
The method of claim 1, wherein said cell is in an in vitro sample.
Claim 4
Dependent Claim
19 words
The method of claim 1, wherein said cell is in a tissue that is connected to a living mammal.
Claim 5
Dependent Claim
16 words
The method of claim 1, wherein said cell is in tissue taken from a living mammal.
Claim 6
Dependent Claim
13 words
The method of claim 1, wherein said cell is in a living mammal.
Claim 7
Dependent Claim
16 words
The method of claim 1, wherein said sealed enclosure encloses an entire mammal comprising said cell.
Claim 8
Dependent Claim
12 words
The method of claim 1, wherein said cell is in endothelial tissue.
Claim 9
Dependent Claim
12 words
The method of claim 1, wherein said cell is in cardiovascular tissue.
Claim 10
Dependent Claim
14 words
The method of claim 1, wherein said nucleic acid molecule is present in solution.
Claim 11
Dependent Claim
19 words
The method of claim 1, wherein said liquid medium is brought into contact with said cell using a catheter.
Claim 12
Independent Claim
57 words
An apparatus for delivering a nucleic acid into a cell, said apparatus comprising: a means for establishing a sealed enclosure enclosing said cell and an extracellular environment of said cell; and a means for establishing an incubation pressure within said sealed enclosure, for facilitating an uptake of said nucleic acid from said extracellular environment into said cell.
Claim 13
Independent Claim
57 words
An apparatus for delivering a nucleic acid into a cell, said apparatus comprising: a means for establishing a sealed enclosure enclosing said cell and an extracellular environment of said cell; and a means for establishing an incubation pressure within said sealed enclosure, for facilitating an uptake of said nucleic acid from said extracellular environment into said cell.
Claim 14
Dependent Claim
24 words
The apparatus of claim 11, wherein said means for establishing said sealed enclosure comprises a protective means for protecting said tissue from experiencing trauma.
Claim 15
Dependent Claim
20 words
The apparatus of claim 12, wherein said protective means comprises an inelastic sheath adapted to be placed around said tissue.
Claim 16
Dependent Claim
23 words
The apparatus of claim 13, wherein said sheath is impermeable, for holding a fluid in a space between said sheath and said tissue.
Claim 17
Dependent Claim
20 words
The apparatus of claim 14, further comprising a sealing means for preventing a leakage of said fluid from said space.
Claim 18
Dependent Claim
35 words
The apparatus of claim 15, further comprising a flushing means for evacuating a fluid from a space contained within a tissue prior to a delivery of said nucleic acid, wherein said tissue comprises said cell.
Claim 19
Dependent Claim
21 words
The apparatus of claim 16, wherein said flushing means comprises a pumping means for passing a flushing gas through said space.
Claim 20
Dependent Claim
21 words
The apparatus of claim 17, further comprising a means for delivering a nucleic acid to an extracellular environment of said cell.
Claim 21
Dependent Claim
17 words
The apparatus of claim 18, wherein said means for delivering said nucleic acid comprises a catheter system.
Claim 22
Dependent Claim
23 words
The apparatus of claim 19, wherein said catheter system comprises two occlusion means for defining said sealed enclosure between said two occlusion means.
Claim 23
Dependent Claim
17 words
The apparatus of claim 11, wherein said means for establishing said sealed enclosure comprises a pressurization chamber.
Claim 24
Independent Claim
47 words
An apparatus for delivering a nucleic acid into a cell, comprising: a means for delivering said nucleic acid to an extracellular environment of said cell; and a means for establishing an incubation pressure around said cell, thereby facilitating an uptake of said nucleic acid into said cell.
Claim 25
Independent Claim
47 words
An apparatus for delivering a nucleic acid into a cell, comprising: a means for delivering said nucleic acid to an extracellular environment of said cell; and a means for establishing an incubation pressure around said cell, thereby facilitating an uptake of said nucleic acid into said cell.
References Cited (Backward Citations)
US5328470A
Cited
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