Seed Promoters

Summary

PLEASE NOTE that patent claims drawn to promoters that are active constitutively in plants, promoters that are active in “fruits” or in “reproductive parts” may be construed to cover seed-preferred promoters. Patent claims to particular sequences or to promoters that drive particular seed-specific genes in certain species (such as phaseolin or napin) may also be applicable, even though they may not mention the word “seed”.

Thus, the list below mentioning a few patents claiming promoters that are broadly seed-specific is NOT a comprehensive list of promoters that are covered by patent claims.

Calgene, Sapporo Breweries and the University of California have filings drawn to seed-specific promoters in broad terms, listed below.

A patent family with several patents granted to Calgene, directed to a transcription cassette having a seed-specific promoter, is noted below, and see also the fruit-specific claims of Calgene elsewhere in this landscape, which may be construed to apply to seeds.

Patents and patent applications that were assigned to Calgene may now be held by Monsanto, to which any inquiries about licensing should be directed.

The granted United States patents US 5420034 and US 5608152 are directed to promoters isolated from specific seed genes (i.e. napin gene) and plants (i.e. Brassica). There are three other granted patents in this patent family, and patent applications in the same patent family may still be pending.

Note that patent claims are not granted the same way in every country, and this patent family presents a perfect example of that. The Australian granted patent claims are broad:

A seed comprising a transcription cassette containing:
- a seed-specific transcriptional initiation region;
- a sequence of interest other than the native sequence regulated by the transcriptional region; and
- a transcriptional termination region.
A transcription construct comprising a polylinker with at least two restriction sites for the insertion of DNA sequences of interest under the control of a seed-specific promoter.
A method to modify the genotype of a seed by the use of a transcription cassette as described above.

Patent Number

Title, Independent Claims and Summary

Assignee

EP 255378 B2

Earliest priority – 31 July 1986
Filed – 30 July 1987
Granted – 13 April 1994
Expected expiry – 30 July 2007

Title – Seed-specific transcriptional regulation

Claim 1
A DNA construct comprising in the 5′ to 3′ direction of transcription:
a napin transcriptional initiation region, joined to a DNA sequence of interest other than
(i) a DNA sequence encoding napin or
(ii) a DNA sequence encoding a mammalian protein or peptide or mammalian viral pathogen peptide or protein; and
a transcription termination region.

Claim 6
A method comprising the production of a DNA construct comprising in the 5′ to 3′ direction of transcription:
a napin transcriptional initiation region wherein said napin transcriptional initiation region is free from the native DNA sequence under the regulatory control of said initiation region, joined to a cloning site, anda transcriptional termination region; provided that said construct does not comprise a DNA sequence encoding a mammalian peptide or protein or mammalian viral pathogen peptide or protein operably linked to said napin transcriptional initiation region.

Claim 8
A method of modifying the genotype of a plant to impart a desired characteristic to seed as distinct from other plant said method comprising:
transforming a host plant cell under genomic integration conditions with a DNA construct comprising in the 5′ to 3′ direction of transcription:
a seed specific napin transcriptional initiation region, joined to a DNA sequence of interest other than (a) a DNA sequence encoding a napin or (b) a DNA sequence encoding a mammalian peptide or protein or a mammalian viral pathogen peptide or protein; and
a transcriptional termination region; and growing said plant to produce seed.

Claim 9
A method for specifically modifying the phenotype of seed as distinct from other plant tissue, said method comprising:
(i) transforming a host plant cell under genomic integration conditions with a DNA construct comprising in the 5′ to 3′ direction of transcription:
a seed specific napin transcriptional initiation region, joined to a DNA sequence of interest other than (a) a DNA sequence encoding napin or (b) a DNA sequence encoding a mammalian peptide or protein or a mammalian viral pathogen peptide or protein; and
a transcriptional termination region; and
(ii) growing a plant under conditions to produce seed, said plant being comprised of cells capable of developingseed tissue and said cells having integrated in their genome said DNA construct.

Claim 10
A method comprising the production of a DNA construct comprising in the 5′ to 3′ direction of transcription:
a seed specific transcriptional initiation region which is from other than the bean phaseolin promoter;
a DNA sequence other than the natural coding sequence joined to said initiation region, wherein said sequence encodes an acyl carrier protein; and
a transcriptional termination region.

Claim12
A method for modifying the genotype of a plant to impart a desired characteristic to seed as distinct from other plant tissues said method comprising:
transforming a host plant cell under genomic integration conditions with a DNA construct comprising in the 5′ to 3′ direction of transcription:
a seed specific transcriptional initiation region;
a DNA sequence encoding an acyl carrier protein joined to said initiation region; and
a transcriptional termination region; and
growing said plant to produce seed.

Claim 13
A method for specifically modifying the phenotype of seed as distinct from other plant tissue, said method comprising:
(i) transforming a host plant cell under genomic integration conditions with a DNA construct comprising in the 5′ to 3′ direction of transcription:
a seed specific transcriptional initiation region;
a DNA sequence encoding an acyl carrier protein joined to said initiation region; and
a transcriptional termination region; and
(ii) growing a plant under conditions to produce seed, said plant being comprised of cells capable of developing seed tissue and said cells having integrated in their genome said DNA construct.

Calgene Inc.

US 5420034

Earliest priority – 31 July 1986
Filed – 8 August 1991
Granted – 30 May 1995
Expected expiry – 30 May 2012

Title – Seed-specific transcriptional regulation

Claim 1
A DNA construct comprising: in the 5′ to 3′ direction of transcription,a transcriptional initiation region from a gene which encodes a product preferentially expressed in a plant seed cell as compared with other plant cells,
a DNA sequence of interest other than the native coding sequence of said gene, and
a transcriptional termination region, wherein said gene is a napin gene, an acyl carrier protein gene or an EA9 gene.

Claim 8
An expression cassette comprising: in the 5′-3′ direction of transcription,a seed-specific transcriptional initiation region wherein said transcriptional initiation region is free from the native DNA sequence under the regulatory control of said initiation region,
a cloning site, and
a transcriptional termination region, wherein said transcriptional initiation region is from a napin gene, an acyl carrier protein gene or an EA9 gene.

Claim 9
An expression cassette comprising: in the 5′-3′ direction of transcription,a transcriptional initiation region and ribosome binding site from a gene expressed in a seed embryo or a seed coat cell or from a gene encoding a seed storage protein,
a linker or polylinker having one or a plurality of restriction sites for insertion of a gene to be expressed under transcriptional control of said transcriptional initiation region, and
a transcriptional termination region, wherein said transcriptional initiation region and said ribosome binding site are from a napin gene, an acyl carrier protein gene or an EA9 gene.

US 5608152

Earliest priority – 31 July 1986
Filed – 30 May 1995
Granted – 4 March 1997
Expected expiry – 4 March 2014

Title – Seed-specific transcriptional regulation

Claim 1
A Brassica plant comprising: a DNA construct comprising, in the 5′ to 3′ direction of transcription,
a transcriptional initiation region from a gene that encodes a product preferentially expressed in a plant seed cell as compared to other plant cells,
a DNA sequence of interest other than the native coding sequence of said gene, and
a transcriptional termination region, wherein said gene is a napin gene, an acyl carrier protein gene or an EA9 gene.

Claim 8
A Brassica seed comprising: a DNA construct comprising, in the 5′ to 3′ direction of transcription,
a transcriptional initiation region from a gene that encodes a product preferentially expressed in a plant seed cell as compared to other plant cells,
a DNA sequence of interest other than the native coding sequence of said gene, and
a transcriptional termination region, wherein said gene is a napin gene, an acyl carrier protein gene or an EA9 gene.

This patent is a Division of US 5420034.

Remarks

Related patents granted in Australia (AU 612326 B2) and New Zealand (NZ 221259). Related application filed in China (CN 87106120) has been withdrawn.

Note: Patent information was last updated on 15 May 2006. Search terms: “seed” in abstract and “Calgene” in applicant. Patent database: PatentLens in combination with INPADOC.

Sapporo Breweries’ patents and applications

The granted US and Australian patents claim an isolated barley β-amylase promoter sequence. However, the independent claims as filed in the European and Canadian patent applications are very broad as they recite a promoter capable of expressing an introduced gene in plant seeds. There is no limitation in the gene source of the seed promoter. In dependent claims the promoter is derived from a beta-amylase gene from barley. It remains to be seen whether the independent claims will be granted as filed.

Patent Number

Title, Independent Claims and Summary

Assignee

US 5952489

Earliest priority – 5 May 1995
Filed – 4 March 1997
Granted – 14 September 1999
Expected expiry – 4 March 2017

Title – Tissue-specific promoter

Claim 1
An isolated barley β-amylase promoter comprising SEQ ID NO: 1.

SapporoBreweries Ltd.

AU 717055 B2

Earliest priority – 5 May 1995
Filed – 4 March 1997
Granted – 14 September 1999
Expected expiry – 4 March 2017

Title – Tissue-specific promoter

Claim 1
An isolated barley β-amylase promoter comprising a nucleic acid
sequence of SEQ ID NO: 1, or a nucleic acid sequence of SEQ ID NO: 1 in which one or more bases are deleted, substituted or added to said sequence and which has promoter activity in plant seeds.

EP 781849 A1

Earliest priority – 5 May 1995
Filed – 5 July 1996
Granted – Pending
Expected expiry – N/A

Title – Tissue-specific promoter

Claim 1
A promoter capable of expressing an introduced gene in plant seeds

Remarks

The related patent application in Canada (CA 2199158) is also pending.

Note: Patent information was last updated on 15 May 2006. Search terms: “seed” in abstract and “Sapporo” in applicant. Patent database: PatentLens and esp@cenet in combination with INPADOC.

University of California‘s patents and applications

Only independent claim 1 as filed of the European patent application is relevant for the present paper as it describes in general terms a recombinant nucleic acid molecule having a seed-maturation specific promoter. The promoter drives the expression of a protein in a subcellular compartment due to the presence of a signal peptide that targets the polypeptide to an intracellular body.

The other independent claims are more specific and describe promoter and signal sequences derived from barley hordein storage protein. It is important to note that claims from pending applications may still be granted.

Patent Number

Title, Independent Claims and Summary

Assignee

US 6642437

Earliest priority – 30 September 1997
Filed – 30 September 1998
Granted – 4 November 2003
Expected expiry – 30 September 2018

Title – Production of proteins in plant seeds

Claim 1
A method for producing seeds containing a selected heterologous protein which is not a seed-storage protein, comprising the steps of:(a) stably transforming monocot plant cells with a chimeric gene having:
(i) a transcriptional regulatory region from the gene of a maturation specific monocot storage protein selected from the group consisting of rice glutelins, rice oryzins, rice prolamines, barley hordeins, oat glutelins, and sorghum kafirins, millet pennisetins, and rye secalins,
(ii) operably linked to said transcriptional regulatory region, a first DNA sequence encoding a monocot seed-specific N-terminal leader sequence capable of targeting a linked polypeptide to a protein storage body in monocot seeds, and
(iii) a second DNA sequence encoding such selected non-seed-storage heterologous protein, and linked in translation frame with the first sequence, such that the first and second sequences encode a fusion protein composed of the selected heterologous non-seed-storage protein and an N-terminal leader sequence,
(b) cultivating plants containing the transformed plant cells under seed-maturation conditions, wherein the expression of the non-seed storage heterologous protein is at least twice that observed with an equivalent chimeric gene lacking the second DNA sequence encoding a monocot seed-specific N-terminal leader sequence, and
(c) harvesting seeds from the cultivated plants.

The Regents of the University of California

AU 746032 B2

Earliest priority – 30 September 1997
Filed – 30 September 1998
Granted – 11 April 2002
Expected expiry – 30 September 2017

Title – Production of proteins in plant seeds

Claim 1
A method of producing a selected heterologous protein which is not a seed-storage protein, comprising the steps of:
(a) stably transforming a monocot plant cell with a chimeric gene comprising:
(i) a transcriptional regulatory region from the gene of a maturation specific monocot storage protein selected from the group consisting of rice glutelins, oryzins, and prolamines, barley hordeins, wheat gliadins and glutenins, maize zeins and glutelines, oat glutelins, and sorghum kafirins, millet pennisetins, and rye secaliiis,
(ii) operably linked to said transcriptional regulatory region, a first DNA sequence encoding a monocot seed-specific leader sequence capable of targeting a linked polypeptide to a protein storage body in monocot seeds, and
(iii) a second DNA sequence encoding the selected heterologous protein, and linked in translation frame with the first sequence, such that the first and second sequences encode a fusion protein composed of the selected protein and an N-terminal leader sequence, and
(b) cultivating plants containing the cell under seed-maturation conditions to produce the selected heterologous protein.

Claim 5
A method of producing a transformed plant comprising the steps of:
a) stably transforming a monocot plant cell with a chimeric gene comprising:
(i) a transcriptional regulatory region from the gene of a maturation specific monocot storage protein selected from the group consisting of rice glutelins, oryzins, and prolamines, barley hordeins, wheat gliadins and glutenins, maize zeins and glutelines, oat glutelins, and sorghum kafirins, millet pennisetins, and rye secalins,
(ii) operably linked to said transcriptional regulatory region, a first DNA sequence encoding a monocot seed-specific leader sequence capable of targeting a linked polypeptide to a protein storage body in monocot seeds, and
(iii) a second DNA sequence encoding a selected heterologous protein which is not a seed-storage protein, and linked in translation frame with the first sequence, such that the first and second sequences encode a fusion protein composed of the selected protein and an N-terminal leader sequence; and
(b) cultivating a plant containing the cell.

EP 1019517 A2

Earliest priority – 30 September 1997
Filed – 30 September 1998
Granted – Pending
Expected expiry – N/A

Title – Production of proteins in plant seeds

Claim 1
A recombinant nucleic acid molecule having a structure P-X or P-SS-X, wherein X is a nucleic acid molecule encoding a polypeptide, P is a seed maturation-specific promoter, and SS is a signal sequence that targets a linked polypeptide to an intracellular body.

Claim 12
A recombinant nucleic acid molecule having a structure Ph-hSS-X, wherein Ph is a hordein promoter, hSS is a hordein signal sequence, and X is a nucleic acid molecule encoding a polypeptide, and where Ph, hSS and X are operably inked.

Claim 30
A method of producing a stably transformed monocotyledenous plant expressing a selected polypeptide in seeds of the plant,comprising:
(a) placing an immature zygotic embryo of the plant on plant growth medium comprising maltose as a sugar source an auxin at a concentration of about 0.1 mg/L to about 5 mg/L, a cytokinin at a concentration of 0 mg/L to about 5 mg/L and copper at a concentration of about 0.1uM to about 50uM, and incubating in dim light conditions so as to form green regenerative tissue;
(b) introducing a nucleic acid molecule into the tissue to produce transformed tissue, wherein the nucleic acid molecule has a structure Ph-hSS-X, wherein Ph is a hordein promoter, hSS is a hordein signal sequence, and X is a nucleic acid molecule encoding the selected polypeptide, and where Ph, hSS and X are operably linked;
(c) incubating the transformed tissue on the plant growth medium such that green structures are observed on the transformed material;
(d) regenerating at least one transformed plant from the green structures; and
(f) growing the transformed plant to produce seed.

Claim 35
A method of producing a stably transformed monocotyledenous plant expressing a selected polypeptide in seeds of the plant,comprising:
(a) placing an immature zygotic embryo of the plant on plant growth medium comprising maltose as a sugar source, an auxin at a concentration of about 0.1 mg/L to about 5 mg/L, a cytokinin at a concentration of0 mg/L to about 5 mg/L and copper at a concentration of about0.1 uM to about 50ltM, and incubating in dim light conditions so as to form green regenerative tissue;
(b) introducing a nucleic acid molecule into the tissue by to produce transformed tissue, wherein the nucleic acid molecule has a structure Ph-X, wherein Ph is a hordein promoter and X is a nucleic acid molecule encoding the selected polypeptide, and where Ph and X are operably linked;
(c) incubating the transformed tissue on the plant growth medium such that green structures are observed on the transformed material;
(d) regenerating at least one transformed plant from the green structures; and
(f) growing the transformed plant to produce seed.

Claim 39
A method of producing a stably transformed monocotyledenous plant expressing a selected polypeptide in seeds of the plant,comprising:
(a) placing an immature zygotic embryo of the plant on plant growth medium comprising maltose as a sugar source, an auxin at a concentration of about 0.1 mg/L to about 5 mg/L, a cytokinin at a concentration of0 mg/L to about 5 mg/L and copper at a concentration of about 0. 11M to about 50I1M, and incubating in dim light conditions so as to form green regenerative tissue;
(b) introducing a nucleic acid molecule into the tissue by to produce transformed tissue, wherein the nucleic acid molecule has a structure P – X or P – SS – wherein X is a nucleic acid molecule encoding a polypeptide, P is a seed maturation-specific promoter, and SS is a signal sequence that targets a linked polypeptide to an intracellular body;
(c) incubating the transformed tissue on the plant growth medium such that green structures are observed on the transformed material;
(d) regenerating at least one transformed plant from the green structures; and
(f) growing the transformed plant to produce seed.

Remarks

An application as a Continuation of US 6642437, which has exactly the same claims as (EP 1019517), is pending in the United States (US 2004/88754 A1). Related applications are also pending in Canada (CA 2305628) and Japan (JP 2001518305)

Note: Patent information was last updated on 15 May 2006. Search terms: “seed” in abstract and “University of California” in applicant. Patent database: PatentLens and esp@cenet in combination with INPADOC.

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Seed Promoters

Summary