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Promoters based on estrogen receptor (ER)

The Rockefeller University has three granted patents in the United States and one granted patent in Australia directed to methods for transforming plant cells with a vector comprising a regulatory region from an ER. It also filed two European patent applications and one Canadian  patent application.

Specific Patent Information

Patent Number

Title, Independent Claims and Summary of Claims

 Assignee
US 6063985

  • Earliest priority – 28 January 1998
  • Filed – 28 January 1998
  • Granted – 16 May 2000
  • Expected expiry – 28 January 2018
 Title – Chemical inducible promoter used to obtain transgenic plants with a silent marker

Claim 1
A method for selecting transgenic plants comprising a silent selectable marker wherein said method comprises the steps of: a) transforming a plant cell with a vector wherein said vector comprises a gene selected from the group consisting of an ipt gene, a CKI1 gene or a gene from the knotted family, wherein said gene is under the control of an inducible promoter; b) growing said plant cells in the absence of an exogenous plant hormone but in the presence of an exogenous inducer of said inducible promoter; and c) excising shoots which develop, wherein said shoots can grow into transgenic plants when grown in the absence of said inducer.

Independent claim 1 is drawn to:

A method for selecting transgenic plants comprising a silent selectable marker by transforming a plant cell with a vector comprising a silent selectable marker, selecting the plant cell in the presence of an inducer and excising shoots for generating trasgenic plants in the absence of the inducer.  The silent selectable marker gene (an ipt gene, a CKI1 gene or a gene from the knotted family) is under the control of an inducible promoter which can be turned on by applying steroid (one such promoter is based on the glucocorticoid receptor as claimed in the dependent claims).

The Rockefeller University
US 6784340

  • Earliest priority – 28 January 1998
  • Filed – 12 November 1999
  • Granted – 31 August 2004
  • Expected expiry – 12 November 2019
 Title – Chemical inducible promoter used to obtain transgenic plants with a silent marker

This patent is a Continuation in part of US 6063985.

Claim 1
A vector comprising a DNA sequence encoding a transcription factor, having the following elements in the 5′ to 3′ direction, i) a promoter, ii) DNA encoding a DNA binding domain of the bacterial repressor LexA, iii) DNA encoding a transactivating domain of VP16, iv) DNA encoding the regulatory domain of an estrogen receptor.
Claim 11
An isolated nucleic acid encoding a transcription factor, comprising, in the 5′ to 3′ direction, i) a promoter, ii) DNA encoding a DNA binding domain of the bacterial repressor LexA, iii) DNA encoding a transactivating domain of VP16, iv) DNA encoding a regulatory domain of an estrogen receptor.
US 6452068

  • Earliest priority – 28 January 1998
  • Filed – 12 November 1999
  • Granted – 17 September 2002
  • Expected expiry – 12 November 2019
 Title – Chemical inducible promoter used to obtain transgenic plants with a silent marker

This patent is a Continuation in part of US 6063985.

Claim 1
A method for selecting a transgenic plant encoding a silent selectable marker wherein said method comprises the steps of: a) transforming a plant cell with a vector wherein said vector comprises DNA encoding a regulatory region of an estrogen receptor, a DNA-binding domain and a transactivating domain, and further wherein said vector comprises nucleic acid encoding a silent selectable marker which promotes shoot formation, wherein said nucleic acid is under the control of an inducible promoter to which the DNA-binding domain binds in response to an inducer which is estrogen or an estrogen derivative; b) growing said plant cell in the absence of an exogenous plant hormone but in the presence of said inducer of said inducible promoter; c) excising shoots which develop, and d) growing said shoots to produce plants; wherein a plant produced from step (d) is selected as being a transgenic plant.
EP 1232273 A2

  • Earliest priority – 12 November 1999
  • Filed – 13 November 2000
  • Granted – Pending
  • Expected expiry – N/A
 Title – Chemical inducible promoters used to obtain transgenic plants with a silent marker

Claim 1
A method for selecting transgenic plants comprising a silent selectable marker wherein said method comprises the stepsof : a) transforming a plant cell with a vector wherein said vector comprises DNA encoding a regulatory region of an estrogen receptor and further wherein said vector comprises a gene which promotes shoot formation, wherein said gene is under the control of an inducible promoter ; b) growing said plant cells in the absence of a plant hormone but in the presence of an inducer of said inducible promoter ; and c) excising shoots which develop, wherein said shoots can grow into transgenic plants when grown in the absence of said inducer.
Claim 14
A method for inducing plant somatic embryo formation comprising the stepsof : a) transforming a plant cell with a vector encoding a gene which promotes somatic embryogenesis, wherein said gene is under the control of an inducible promoter ; and b) growing said plant cells in the absence of a plant hormone but in the presence of an inducer of said inducible promoter, wherein somatic embryos will develop.
Claim 26
A method for selecting transgenic plants wherein said method comprises growing a transgenic plant, comprising an antibiotic resistance gene under the control of a promoter comprising DNA encoding a regulatory domain of an estrogen receptor inducible by17-P- estradiol or 4-hydroxyl tamoxifen, in the presence of an antibiotic, wherein said antibiotic is one to which resistance is conferred by said antibiotic resistance gene, and in the presence of17-p- estradiol or 4-hydroxyl tamoxifen.
Claim 27
A method for selecting transgenic plants wherein said method comprises growing a transgenic plant, comprising a herbicide resistance gene wherein said herbicide resistance gene is under the control of a promoter comprising DNA encoding a regulatory domain of an estrogen receptor inducible by17-P-estradiol or 4-hydroxyl tamoxifen, in the presence of a herbicide, wherein said herbicide is one to which resistance is conferred by said herbicide resistance gene, and in the presence of17-P-estradiol or 4-hydroxyl tamoxifen.

European application EP 1 232 273 A2 claims methods for selecting transgenic plants by inserting a vector having a gene of interest under the control of an ER. When an inducer compound is added, the transformed plants express the gene of interest. The genes under the control of the inducible promoter are:

  • genes promoting shoot formation;
  • genes promoting somatic embryogenesis;
  • an antibiotic resistance gene; and
  • a herbicide resistance gene.

The compounds 17-beta-estradiol and 4-hydroxyl tamoxifen are cited as inducers of the transcription system.

Although the title of the application reads chimeric promoters, these are not subject matter of the independent claims as filed. (??- It doesn’t according to this table) However, some dependent claims mention the use however of the DNA-binding domain of the bacterial repressorLexA assembled with the regulatory domain of the ER.

Remarks

 An United States application (US 2003/150013 A1) , as a Division of US 6452068, and  a Canadian application (CA 2391003) are pending.
EP 1242604 A2

  • Earliest priority – 12 November 1999
  • Filed – 13 November 2000
  • Granted – Pending
  • Expected expiry – N/A
 Title – Chemical inducible promoters used to obtain transgenic plants with a silent marker and organisms and cells and methods of using same for screening for mutations

Claim 1
A method for selecting transgenic lettuce plants comprising a silent selectable marker wherein said method comprises the stepsof : a) transforming lettuce root cells with a vector wherein said vector comprises a gene selected from the group consisting of an ipt gene, aCKI1 gene, a gene from the knotted family, and a gene the expression of which is capable of promoting shoot regeneration, wherein said gene is under the control of an inducible promoter ; b) growing said lettuce root cells to allow shoot development ; and c) excising shoots which develop from plants having a shooty phenotype, wherein said shoots can grow into normal transgenic plants when grown in the absence of said inducer.
Claim 11
A vector comprising a chemically inducible promoter wherein said vector comprises
DNA encoding an estrogen receptor.
Claim 25
A vector comprising i) a constitutive promoter, ii) DNA encoding a DNA binding domain of bacterial repressor LexA, iii) DNA encoding a transactivating domain of VP16, iv)DNA encoding an estrogen receptor, and v) one or more LexA binding sites.
Claim 26
A nucleic acid comprising a chemically inducible promoter wherein said nucleic acid further comprises DNA encoding an estrogen receptor.
Claim 41
A nucleic acid comprising i) a constitutive promoter, ii) DNA encoding a DNA binding domain of bacterial repressor LexA, iii) DNA encoding a transactivating domain of VP 16, iv) DNA encoding an estrogen receptor, and v) one or more LexA binding sites.
Claim 42
A transgenic lettuce plant or transgenic lettuce plant cell comprising a vector wherein said vector comprises a chemically inducible promoter.
Claim 43
A transgenic plant or transgenic plant cell comprising a vector wherein said vector comprises a chemically inducible promoter which can be induced by an estrogen.
Claim 50
A method for making a transgenic plant display a fluorescent design, a word or words wherein said method comprises the stepsof : a) preparing a transgenic plant which comprises a luciferase gene under the control of a chemically inducible promoter which is controlled by an estrogen ; and b) placing a chemical which induces said chemically inducible promoter onto said transgenic plant in the pattern of the design, word or words which are desired ; whereby said plant will produce luciferase and will fluoresce in the pattern in which the chemically inducible promoter was placed onto said transgenic plant.
Claim 52
A transgenic lettuce plant comprising an antibiotic resistance gene wherein said antibiotic resistance gene is under the control of an inducible promoter.
Claim 53
A transgenic plant comprising an antibiotic resistance gene wherein said antibiotic resistance gene is under the control of an inducible promoter, wherein said inducible promoter comprises DNA encoding a regulatory domain of an estrogen receptor.
Claim 57
A transgenic plant comprising a herbicide resistance gene wherein said herbicide resistance gene is under the control of an inducible promoter, wherein said inducible promoter comprises DNA encoding a regulatory domain of an estrogen receptor.
Claim 61
An organism or a cell comprising a gene wherein a natural promoter of said gene is lacking or inoperative and said gene is under the control of a transgenic inducible promoter.
Claim 74
A method to screen for mutations in a gene of an organism or cell comprising a) preparing an organism or a cell wherein a natural promoter of said gene is lacking or inoperative and said gene is under the control of a transgenic inducible promoter ; and b) growing said organism or cell.

The claims range those drawn to organisms or cells comprising a transgenic inducible promoter to a specific chimeric inducible promoter corresponding to the XVE promoter mentioned above in “scientific aspects” and to lettuce plants transformed with an inducible promoter driving particular genes promoting shoot development and antibiotic resistance.

The broadest claims originally  filed recite:

  • an organism or cell comprising a transgenic inducible promoter (which can be any type of inducible promoter) which controls a gene whose natural promoter is inoperative or lacking;
  • a method for screening mutations in an organism or cell gene by putting that gene under the control of a transgenic inducible promoter;
  • a nucleic acid containing a chemically-inducible promoter having an ER;
  • a vector containing such a chemically-inducible promoter;
  •  transgenic plant or plant cell containing such chemically-inducible promoter.

In other claims, the chimeric inducible promoter comprises:

  • a constitutive promoter;
  • a DNA-binding domain of bacterial repressor LexA;
  • a transactivating domain of VP16;
  • an estrogen receptor; and
  • at least one LexA binding site.

In addition, the claims recite either a transgenic plant cell or specifically a transgenic lettuce plant comprising an inducible promoter, which in some cases explicitly comprises an ER. The same genes disclosed in EP 1282273 A2 under the control of the inducible promoter are in the claims as filed, with the exception of genes promoting somatic embryogenesis.

The Rockefeller University
AU 782958 B2

  • Earliest priority – 12 November 1999
  • Filed – 13 November 2000
  • Granted – 15 September 2005
  • Expected expiry – 13 November 2020
 Title – Chemical inducible promoters used to obtain transgenic plants with a silent marker and organisms and cells and methods of using same for screening for mutations

Claim 1
A vector comprising a nucleic acid encoding a transcription factor, said nucleic acid comprising in the 5′ to 3′ direction, i) a promoter, ii) DNA encoding a DNA binding domain of the bacterial repressor LexA, iii) DNA encoding a transactivating domain of VP16, and iv) DNA encoding the regulatory domain of an estrogen receptor.
Claim 11
A nucleic acid encoding a transcription factor, comprising, in the 5′ to 3′ direction, i) a promoter, ii) DNA encoding a DNA binding domain of the bacterial repressor LexA, iii) DNA encoding a transactivating domain of VP16, and iv) DNA encoding a regulatory domain of an estrogen receptor.

Note: Patent information on this page was last updated on 22 March 2006.