US5107065A
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US
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Anti-sense regulation of gene expression in plant cells
Assignee: Monsanto Co
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Filing: 1988-08-30
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Granted: 1992-04-21
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Expires: 2008-08-30
Abstract
Abstract Regulation of expression of genes encoded for in plant cell genomes is achieved by integration of a gene under the transcriptional control of a promoter which is functional in the host and in which the transcribed strand of DNA is complementary to the strand of DNA that is transcribed from the endogenous gene(s) one wishes to regulate. The integrated gene, referred to as anti-sense, provides an RNA sequence capable of binding to naturally existing RNAs, exemplified by polygalacturonase, and inhibiting their expression, where the anti-sense sequence may bind to the coding, non-coding, or both, portions of the RNA. The antisense construction may be introduced into the plant cells in a variety of ways and be integrated into the plant genome for inducible or constitutive transcription of the anti-sense sequence. A wide variety of plant cell properties may be modifed by employing this technique. The pCGN978xK12 was deposited at the A.T.C.C. on Mar. 25, 1986, and given Accession No. 67064 and pCGN1401 was deposited on Oct. 7, 1986 at the A.T.C.C. and given Accession No. 67227.
Inventors
- 👤 Christine K. Shewmaker
- 👤 Jean C. Kridl
- 👤 William R. Hiatt
- 👤 Vic Knauf
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 19:36
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/11; C12N15/113; C12N15/1137; C12N15/63; C12N15/79; C12N15/82; C12N15/8216; C12N15/8218; C12N9/00; C12N9/14; C12N9/24; C12N9/2402; C12Y; C12Y302/00; C12Y302/01; C12Y302/01015; C12N2310/00; C12N2310/10; C12N2310/11; C12N2310/111
Claims Analysis
Total Claims: 36
Claim 1
Independent Claim
105 words
A method for inhibiting the expression of a gene indigenous to a plant cell, said method comprising: integrating into the genome of a plant cell a construct comprising in the 5'-3' direction of transcription a promoter functional in said plant cell, a dsDNA sequence wherein the transcribed strand of said dsDNA is complementary to RNA indigenous to said cell, and a termination region functional in said cell whereby a transformed cell is obtained; and growing said transformed plant cell, whereby said complementary strand is transcribed and binds to said RNA indigenous to said cell, thereby inhibiting expression of said gene indigenous to said plant cell.
Claim 2
Independent Claim
105 words
A method for inhibiting the expression of a gene indigenous to a plant cell, said method comprising: integrating into the genome of a plant cell a construct comprising in the 5'-3' direction of transcription a promoter functional in said plant cell, a dsDNA sequence wherein the transcribed strand of said dsDNA is complementary to RNA indigenous to said cell, and a termination region functional in said cell whereby a transformed cell is obtained; and growing said transformed plant cell, whereby said complementary strand is transcribed and binds to said RNA indigenous to said cell, thereby inhibiting expression of said gene indigenous to said plant cell.
Claim 3
Dependent Claim
13 words
A method according to claim 1, wherein said indigenous RNA is messenger RNA.
Claim 4
Dependent Claim
23 words
A method according to claim 2, wherein the transcribed strand includes at least 15 nt of the 5'-untranslated sequence of said messenger RNA.
Claim 5
Dependent Claim
23 words
A method according to claim 2, wherein the transcribed strand includes at least 15 nt of the 3'-untranslated sequence of said messenger RNA.
Claim 6
Dependent Claim
16 words
A method according to claim 2, wherein said inhibiting results in alteration of a phenotypic trait.
Claim 7
Dependent Claim
13 words
A method according to claim 1, wherein said plant cell is a monocotyledon.
Claim 8
Dependent Claim
13 words
A method according to claim 1, wherein said plant cell is a dicotyledon.
Claim 9
Dependent Claim
13 words
A method according to claim 1, wherein said plant cell is a gymnosperm.
Claim 10
Independent Claim
109 words
A method for inhibiting expression of an indigenous gene in a plant cell, said method comprising: integrating into the genome of said plant cell a construct comprising in the 5'-3' direction of transcription a promoter functional in said plant cell, a dsDNA sequence wherein the transcribed strand of said dsDNA is complementary to at least 15 nt of the translated region of messenger RNA indigenous to said plant cell and a termination region functional in said cell whereby a transformed cell is obtained; and growing said transformed plant cell, whereby said complementary strand is transcribed and binds to said indigenous messenger RNA and expression of said gene is inhibited.
Claim 11
Independent Claim
109 words
A method for inhibiting expression of an indigenous gene in a plant cell, said method comprising: integrating into the genome of said plant cell a construct comprising in the 5'-3' direction of transcription a promoter functional in said plant cell, a dsDNA sequence wherein the transcribed strand of said dsDNA is complementary to at least 15 nt of the translated region of messenger RNA indigenous to said plant cell and a termination region functional in said cell whereby a transformed cell is obtained; and growing said transformed plant cell, whereby said complementary strand is transcribed and binds to said indigenous messenger RNA and expression of said gene is inhibited.
Claim 12
Dependent Claim
17 words
A method according to claim 9, wherein said construct further comprises at least the right T-DNA border.
Claim 13
Dependent Claim
21 words
A method according to claim 10, wherein said construct further comprises at least one marker for selection of said transformed cells.
Claim 14
Dependent Claim
19 words
A method according to claim 9, wherein said transcribed strand comprises at least 15 nt of the untranslated region.
Claim 15
Dependent Claim
23 words
A plant cell wherein expression of a gene indigenous to said cell is inhibited according to the method of claim 1 or 9.
Claim 16
Dependent Claim
11 words
A plant cell according to claim 13, which is a monocotyledon.
Claim 17
Dependent Claim
11 words
A plant cell according to claim 13, which is a dicotyledon.
Claim 18
Dependent Claim
11 words
A plant cell according to claim 13, which is a gymnosperm.
Claim 19
Dependent Claim
44 words
A DNA construct comprising in the 5'-3' direction of transcription a transcriptional initiation region functional in a plant cell, a dsDNA sequence having as the transcribed strand a sequence complementary to an RNA sequence indigenous to a plant cell, and a transcriptional termination region.
Claim 20
Dependent Claim
44 words
A DNA construct comprising in the 5'-3' direction of transcription a transcriptional initiation region functional in a plant cell, a dsDNA sequence having as the transcribed strand a sequence complementary to an RNA sequence indigenous to a plant cell, and a transcriptional termination region.
Claim 21
Dependent Claim
11 words
A plant cell comprising a DNA construct according to claim 17.
Claim 22
Dependent Claim
11 words
A plant cell according to claim 18, which is a monocotyledon.
Claim 23
Dependent Claim
11 words
A plant cell according to claim 18, which is a dicotyledon.
Claim 24
Dependent Claim
11 words
A plant cell according to claim 18, which is a gymnosperm.
Claim 25
Dependent Claim
26 words
A DNA construct according to claim 17, further comprising a DNA sequence encoding a marker which provides for selection of a plant cell containing said construct.
Claim 26
Dependent Claim
14 words
A DNA construct according to claim 17, further comprising at least one T-DNA border.
Claim 27
Dependent Claim
26 words
A DNA construct according to claim 23, further comprising a DNA sequence encoding a marker which provides for selection of a prokaryotic cell containing said construct.
Claim 28
Dependent Claim
19 words
A plasmid comprising a replication system functional in a prokaryotic host and a DNA construct according to claim 17.
Claim 29
Dependent Claim
17 words
A plasmid comprising a replication system functional in Agrobacterium and a DNA construct according to claim 23.
Claim 30
Independent Claim
104 words
A method for reducing expression of a gene indigenous to a plant cell, said method comprising: integrating into the genome of a plant cell a construct comprising in the 5'-3' direction of transcription a promoter functional in said plant cell, a dsDNA sequence wherein the transcribed strand of said dsDNA is complementary to RNA indigenous to said cell, and a termination region functional in said cell whereby a transformed cell is obtained; and growing said transformed plant cell, whereby said complementary strand is transcribed and binds to said RNA indigenous to said cell, thereby reducing expression of said gene indigenous to said plant cell.
Claim 31
Independent Claim
104 words
A method for reducing expression of a gene indigenous to a plant cell, said method comprising: integrating into the genome of a plant cell a construct comprising in the 5'-3' direction of transcription a promoter functional in said plant cell, a dsDNA sequence wherein the transcribed strand of said dsDNA is complementary to RNA indigenous to said cell, and a termination region functional in said cell whereby a transformed cell is obtained; and growing said transformed plant cell, whereby said complementary strand is transcribed and binds to said RNA indigenous to said cell, thereby reducing expression of said gene indigenous to said plant cell.
Claim 32
Dependent Claim
16 words
A method according to claim 27, wherein said reducing results in alteration of a phenotypic trait.
Claim 33
Independent Claim
108 words
A method for reducing expression of a gene in a plant cell, said method comprising: integrating into the genome of said plant cell a construct comprising in the 5'-3' direction of transcription a promoter functional in said plant cell, a dsDNA sequence wherein the transcribed strand of said dsDNA is complementary to at least 15 nt of the translated region of messenger RNA indigenous to said plant cell, and a termination region functional in said cell whereby a transformed cell is obtained; and growing said transformed plant cell, whereby said complementary strand is transcribed and binds to said indigenous messenger RNA and expression of said gene is reduced.
Claim 34
Independent Claim
108 words
A method for reducing expression of a gene in a plant cell, said method comprising: integrating into the genome of said plant cell a construct comprising in the 5'-3' direction of transcription a promoter functional in said plant cell, a dsDNA sequence wherein the transcribed strand of said dsDNA is complementary to at least 15 nt of the translated region of messenger RNA indigenous to said plant cell, and a termination region functional in said cell whereby a transformed cell is obtained; and growing said transformed plant cell, whereby said complementary strand is transcribed and binds to said indigenous messenger RNA and expression of said gene is reduced.
Claim 35
Dependent Claim
44 words
A plant and progeny thereof derived from a cell having inhibited or reduced expression of a gene indigenous to said cell and produced according to the method of any one of claims 1, 9, 27 or 29, wherein said cell is a dicotyledenous cell.
Claim 36
Dependent Claim
17 words
A dicotyledonous plant and progeny thereof derived from a cell comprising a plasmid according to claim 25.
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