US5578475A
Expired / Lapsed
US
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Composition and methods for transfecting eukaryotic cells
Assignee: Life Technologies Corp
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Filing: 1994-07-12
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Granted: 1996-11-26
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Expires: 2014-07-12
Abstract
Abstract The present invention provides compositions and methods for transfecting eukaryotic cells which comprises a nucleic acid, a cationic lipid capable of forming a complex with said nucleic acid, and a viral agent which is an active or inactive enveloped virus or a component of an enveloped virus.
Inventors
- 👤 Joel A. Jessee
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 11:09
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/005; A; A61; A61K; A61K9/00; A61K9/10; A61K9/127; A61K9/1271; A61K9/1272; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/85; C12N15/86; C12N15/87; C12N15/88; C12N2760/00; C12N2760/00011; C12N2760/16011; C12N2760/16022; C12N2760/20011; C12N2760/20211; C12N2760/20222; C12N2770/00; C12N2770/00011; C12N2770/36011; C12N2770/36111; C12N2770/36141; C12N2770/36143
Claims Analysis
Total Claims: 37
Claim 1
Independent Claim
85 words
A composition for transfecting a eukaryotic cell wherein said composition comprises a nucleic acid, a cationic lipid composition selected from the group consisting of a 3:1 (w/w) mixture of the polycationic lipid, 2,3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propanaminiumtrifluoroacetate and dioleoylphosphatidylethanolamine, and a 1:1 (w/w) mixture of N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammoniumchloride and dioleoylphosphatidylethanolamine and a viral agent which enhances transfection two-fold or more by said cationic lipid composition, which viral agent is an alphavirus or a component of an alphavirus and wherein said nucleic acid is not a nucleic acid of said viral agent.
Claim 2
Dependent Claim
85 words
A composition for transfecting a eukaryotic cell wherein said composition comprises a nucleic acid, a cationic lipid composition selected from the group consisting of a 3:1 (w/w) mixture of the polycationic lipid, 2,3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propanaminiumtrifluoroacetate and dioleoylphosphatidylethanolamine, and a 1:1 (w/w) mixture of N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammoniumchloride and dioleoylphosphatidylethanolamine and a viral agent which enhances transfection two-fold or more by said cationic lipid composition, which viral agent is an alphavirus or a component of an alphavirus and wherein said nucleic acid is not a nucleic acid of said viral agent.
Claim 3
Dependent Claim
24 words
The transfection composition according to claim 1 wherein said cationic lipid composition is a 3:1 (w/w) mixture of the polycationic lipid, 2,3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propanaminiumtrifluoroacetate and dioleoylphosphatidylethanolamine.
Claim 4
Dependent Claim
20 words
The transfection composition according to claim 1 wherein said cationic lipid composition is a 1:1(w/w) mixture of N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammoniumchloride and dioleoylphosphatidylethanolamine.
Claim 5
Dependent Claim
18 words
The transfection composition of claim 1 wherein said alphavirus is a Semliki Forest virus or a Sindbis virus.
Claim 6
Dependent Claim
14 words
The transfection composition of claim 4 wherein said alphavirus is a Semliki Forest virus.
Claim 7
Dependent Claim
15 words
The transfection composition of claim 5 wherein said alphavirus is an inactive Semliki Forest virus.
Claim 8
Dependent Claim
15 words
The transfection composition of claim 6 wherein said Semliki Forest virus is a replication-deficient virus.
Claim 9
Dependent Claim
18 words
The transfection composition of claim 5 wherein said vital agent comprises a spike glycoprotein of Semliki Forest virus.
Claim 10
Dependent Claim
21 words
The transfection composition of claim 5 wherein said viral agent is a multimer of a spike glycoprotein of Semliki Forest virus.
Claim 11
Dependent Claim
21 words
The transfection composition of claim 5 wherein said vital agent is a peptide of a spike glycoprotein of Semliki Forest virus.
Claim 12
Dependent Claim
15 words
The transfection composition of claim 6 wherein the alphavirus is a heat-inactivated Semliki Forest virus.
Claim 13
Dependent Claim
13 words
The transfection composition of claim 1 wherein said alphavirus is an inactive alphavirus.
Claim 14
Dependent Claim
19 words
The transfection composition of claim 1 wherein said cationic lipid composition is formed into a cationic liposome or vesicle.
Claim 15
Dependent Claim
18 words
The transfection composition of claim 13 wherein said viral agent is incorporated into said cationic liposome or vesicle.
Claim 16
Dependent Claim
17 words
The transfection composition of claim 13 wherein said cationic lipid composition is fused with said viral agent.
Claim 17
Dependent Claim
17 words
The transfection composition of claim 1 wherein said viral agent is a Semliki Forest virus spike glycoprotein.
Claim 18
Dependent Claim
16 words
The transfection composition of claim 13 wherein said liposome or vesicle further comprises a 3β-OH sterol.
Claim 19
Dependent Claim
23 words
A method for transfecting a eukaryotic cell with a nucleic acid which comprises contacting said cell with the transfection composition of claim 1.
Claim 20
Dependent Claim
18 words
The method of claim 18 wherein said viral agent in said transfection composition is a Semliki Forest virus.
Claim 21
Dependent Claim
19 words
The method of claim 18 wherein said viral agent in said transfection composition is an inactive Semliki Forest virus.
Claim 22
Dependent Claim
22 words
The method of claim 18 wherein said vital agent in said transfection composition is a spike glycoprotein of a Semliki Forest virus.
Claim 23
Dependent Claim
26 words
The method of claim 18 wherein the cationic lipid composition in said transfection composition is a 3:1 (w/w) mixture of the polycationic lipid, 2,3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propanaminiumtrifluoroacetate and dioleoylphosphatidylethanolamine.
Claim 24
Dependent Claim
23 words
The method of claim 18 wherein the cationic lipid composition of said transfection composition is a 1:1(w/w) mixture of N-[1- (2,3-dioleyloxy)propyl]-N,N,N-trimethylammoniumchloride and dioleoylphosphatidylethanolamine.
Claim 25
Dependent Claim
22 words
The method of claim 18 wherein said cationic lipid composition in said transfection composition is formed into a cationic liposome or vesicle.
Claim 26
Dependent Claim
21 words
The method of claim 24 wherein said vital agent of said transfection composition is incorporated into said cationic liposome or vesicle.
Claim 27
Dependent Claim
20 words
The method of claim 24 wherein said cationic lipid composition of said transfection composition is fused with said vital agent.
Claim 28
Dependent Claim
16 words
The method of claim 25, wherein said cationic liposome or vesicle further comprises a 3β-OH sterol.
Claim 29
Dependent Claim
35 words
The method of claim 18 which further comprises the step of introducing a 3β-OH sterol into the cell membrane of said eukaryotic cell prior to or simultaneously with contacting said cell with said transfection composition.
Claim 30
Dependent Claim
11 words
The method of claim 28 wherein said 3β-OH sterol is cholesterol.
Claim 31
Dependent Claim
24 words
The method of claim 28 wherein said 3β-OH sterol is introduced into the cell membrane of said eukaryotic cell by lipid fusion or exchange.
Claim 32
Dependent Claim
33 words
The method of claim 28 wherein said 3β-OH sterol is introduced into the cell membrane of said eukaryotic cell by growing or incubating said cells in a medium rich in said 3β-OH sterol.
Claim 33
Dependent Claim
13 words
The method of claim 18 wherein said eukaryotic cell is a suspension cell.
Claim 34
Dependent Claim
13 words
The method of claim 18 wherein said eukaryotic cell is an animal cell.
Claim 35
Dependent Claim
25 words
The method of claim 18 wherein said eukaryotic cell is selected from the group consisting of animal primary cells and animal embryo stem line cells.
Claim 36
Dependent Claim
21 words
The method of claim 18 wherein said eukaryotic cell is selected from the group consisting of human fibroblasts, keratinocytes and macrophages.
Claim 37
Dependent Claim
16 words
The transfection composition of claim 1 wherein said component of an alphavirus is a spike glycoprotein.
References Cited (Backward Citations)
US4946787A
Cited
WO1991016024A1
Cited
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| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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