US4959317A
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Site-specific recombination of DNA in eukaryotic cells
Assignee: EIDP Inc
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Filing: 1987-04-29
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Granted: 1990-09-25
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Expires: 2007-04-29
Abstract
Abstract A method for producing site-specific recombination of DNA in eukaryotic cells at regions designated lox sites is disclosed.
Inventors
- 👤 Brian L. Sauer
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 22:05
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/87; C12N15/90; C12N15/902; C12N15/907; C12N15/63; C12N15/79; C12N15/80; C12N15/81
Claims Analysis
Total Claims: 49
Claim 1
Independent Claim
50 words
A method for producing site-specific recombination of DNA in eukaryotic cells, comprising, (a) introducing into the cells a first DNA sequence comprising a first lox site and a second DNA sequence comprising a second lox site, and (b) contacting the lox sites with Cre, thereby producing the site specific recombination.
Claim 2
Independent Claim
50 words
A method for producing site-specific recombination of DNA in eukaryotic cells, comprising, (a) introducing into the cells a first DNA sequence comprising a first lox site and a second DNA sequence comprising a second lox site, and (b) contacting the lox sites with Cre, thereby producing the site specific recombination.
Claim 3
Dependent Claim
22 words
A method as defined in claim 1, wherein a third DNA sequence comprising a cre gene is also introduced into the cells.
Claim 4
Dependent Claim
32 words
A method as defined in claim 2, wherein the third DNA sequence further comprises a regulatory nucleotide sequence and expression of the cre gene is produced by activating the regulatory nucleotide sequence.
Claim 5
Dependent Claim
25 words
A method as defined in claim 3, wherein the first and second DNA sequences are introduced into the cells connected by a pre-selected DNA segment.
Claim 6
Dependent Claim
33 words
A method as defined in claim 4, wherein the first and second lox sites have the same orientation and the site specific recombination of DNA is a deletion of the pre-selected DNA segment.
Claim 7
Dependent Claim
16 words
A method as defined in claim 5, wherein the cre gene is isolated from bacteriophage P1.
Claim 8
Dependent Claim
18 words
A method as defined in claim 5, wherein the first and second lox sites are loxP or loxC2.
Claim 9
Dependent Claim
25 words
A method as defined in claim 7, wherein the pre-selected DNA segment is a gene for a structural protein, an enzyme, or a regulatory molecule.
Claim 10
Dependent Claim
15 words
A method as defined in claim 8, wherein the eukaryotic cell is a mammalian cell.
Claim 11
Dependent Claim
13 words
A method as defined in claim 8, wherein the eukaryotic cell is yeast.
Claim 12
Dependent Claim
33 words
A method as defined in claim 4, wherein the first and second lox sites have opposite orientations and the site-specific recombination is an inversion of the nucleotide sequence of the pre-selected DNA segment.
Claim 13
Dependent Claim
16 words
A method as defined in claim 11, wherein the cre gene is isolated from bacteriophage P1.
Claim 14
Dependent Claim
18 words
A method as defined in claim 12, wherein the first and second lox sites are loxP or loxC2.
Claim 15
Dependent Claim
25 words
A method as defined in claim 13, wherein the pre-selected DNA segment is a gene for a structural protein, an enzyme, or a regulatory molecule.
Claim 16
Dependent Claim
37 words
A method as defined in claim 1, wherein the second and third DNA sequences are introduced into two different DNA molecules and the site-specific recombination is a reciprocal exchange of DNA segments proximate to the lox sites.
Claim 17
Dependent Claim
16 words
A method as defined in claim 15, wherein the cre gene is isolated from bacteriophage P1.
Claim 18
Dependent Claim
18 words
A method as defined in claim 16, wherein the first and second lox sites are loxP and loxC2.
Claim 19
Dependent Claim
12 words
A eukaryotic cell transformed with a DNA sequence comprising a cre gene.
Claim 20
Dependent Claim
12 words
A eukaryotic cell transformed with a DNA sequence comprising a cre gene.
Claim 21
Dependent Claim
12 words
A eukaryotic cell transformed with a DNA sequence comprising a lox site.
Claim 22
Dependent Claim
12 words
A eukaryotic cell transformed with a DNA sequence comprising a lox site.
Claim 23
Independent Claim
71 words
A method for producing site-specific recombination of DNA in yeast, comprising: (a) introducing into the DNA the following DNA sequences: (i) a DNA sequence comprising a regulatory nucleotide sequence and a cre gene, (ii) a DNA sequence comprising a first lox site, and (iii) a DNA sequence comprising a second lox site, and (b) activating the regulatory nucleotide sequence thereby effecting expression of the cre gene and producing the site-specific recombination.
Claim 24
Independent Claim
71 words
A method for producing site-specific recombination of DNA in yeast, comprising: (a) introducing into the DNA the following DNA sequences: (i) a DNA sequence comprising a regulatory nucleotide sequence and a cre gene, (ii) a DNA sequence comprising a first lox site, and (iii) a DNA sequence comprising a second lox site, and (b) activating the regulatory nucleotide sequence thereby effecting expression of the cre gene and producing the site-specific recombination.
Claim 25
Dependent Claim
27 words
A method as defined in claim 1, wherein the DNA sequences comprising lox sites are introduced into the DNA in yeast connected by a pre-selected DNA segment.
Claim 26
Dependent Claim
16 words
A method as defined in claim 21, wherein the cre gene is isolated from bacteriophage P1.
Claim 27
Dependent Claim
19 words
A method as defined in claim 22, wherein the fir t and second lox sites are loxP or loxC2.
Claim 28
Dependent Claim
42 words
A strain of yeast transformed with the following DNA sequences: (i) a DNA sequence comprising a regulatory nucleotide sequence and a cre gene, (ii) a DNA sequence comprising a first lox site, and (iii) a DNA sequence comprising a second lox site.
Claim 29
Dependent Claim
42 words
A strain of yeast transformed with the following DNA sequences: (i) a DNA sequence comprising a regulatory nucleotide sequence and a cre gene, (ii) a DNA sequence comprising a first lox site, and (iii) a DNA sequence comprising a second lox site.
Claim 30
Dependent Claim
23 words
A strain of yeast as defined in claim 24, wherein the DNA sequences comprising lox sites are connected by a pre-selected DNA segment.
Claim 31
Dependent Claim
18 words
A strain of yeast as defined in claim 25, wherein the cre gene is isolated from bacteriophage P1.
Claim 32
Dependent Claim
20 words
A strain of yeast as defined in claim 26, wherein the first and second lox sites are loxP or loxC2.
Claim 33
Dependent Claim
9 words
Yeast strain BSY90 having deposit accession number ATCC 20772.
Claim 34
Dependent Claim
9 words
Yeast strain BSY90 having deposit accession number ATCC 20772.
Claim 35
Dependent Claim
9 words
Yeast strain pBSY23 having deposit accession number ATCC 20773.
Claim 36
Dependent Claim
9 words
Yeast strain pBSY23 having deposit accession number ATCC 20773.
Claim 37
Dependent Claim
20 words
A plasmid having a regulatory nucleotide sequence and a cre gene, said plasmid being capable of transforming a eukaryotic cell.
Claim 38
Dependent Claim
20 words
A plasmid having a regulatory nucleotide sequence and a cre gene, said plasmid being capable of transforming a eukaryotic cell.
Claim 39
Dependent Claim
16 words
A plasmid as defined in claim 30, wherein the cre gene is isolated from bacteriophage P1.
Claim 40
Dependent Claim
22 words
A plasmid as defined in claim 30, wherein the plasmid is pBS39 characterized by the restriction enzyme map shown in FIG. 1.
Claim 41
Dependent Claim
22 words
A plasmid as defined in claim 30, wherein the plasmid is pBS49 characterized by the restriction enzyme map shown in FIG. 1.
Claim 42
Dependent Claim
17 words
A plasmid having at least one lox site, said plasmid being capable of transforming a eukaryotic cell.
Claim 43
Dependent Claim
17 words
A plasmid having at least one lox site, said plasmid being capable of transforming a eukaryotic cell.
Claim 44
Dependent Claim
24 words
A plasmid as defined in claim 34, wherein a first lox site and a second lox site are connected by a pre-selected DNA segment.
Claim 45
Dependent Claim
17 words
A plasmid as defined in claim 35, wherein the first and second lox sites are LoxP sites.
Claim 46
Dependent Claim
49 words
A plasmid as defined in claim 34, wherein the plasmid is pBS44 characterized by the restriction enzyme map shown in FIG. 6, or a derivative thereof carrying a pre-selected DNA segment other than or in addition to the LEU2 gene and located between the first and second lox sites.
Claim 47
Dependent Claim
49 words
A plasmid as defined in claim 34 wherein the plasmid is pBS47 characterized by the restriction enzyme map shown in FIG. 6, or a derivative thereof carrying a pre-selected DNA segment other than or in addition to the LEU2 gene and located between the first and second lox sites.
Claim 48
Dependent Claim
49 words
A plasmid as defined in claim 34 wherein the plasmid is pBS42 characterized by the restriction enzyme map shown in FIG. 2, or a derivative thereof carrying a pre-selected DNA segment other than or in addition to the LEU2 gene and located between the first and second lox site.
Claim 49
Dependent Claim
49 words
A plasmid as defined in claim 34 wherein the plasmid s pBS43 characterized by the restriction enzyme map shown in FIG. 2, or a derivative thereof carrying a pre-selected DNA segment other than or in addition to the LEU2 gene and located between the first and second lox sites.
References Cited (Backward Citations)
US4626505A
Cited
US4673640A
Cited
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| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1993-09-24 | 1994-03-26 | 1994-09-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1997-09-24 | 1998-03-26 | 1998-09-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2001-09-24 | 2002-03-26 | 2002-09-25 | $7700.00 | Window Closed / Lapsed |