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Patents granted to Syngenta Mogen B.V.

The following United States and EP patents assigned to Syngenta Mogen B.V. are directed to the insertion of specific sequences into a target locus of a plant genome through homologous recombination. The gene of interest and the homologous sequences are part of a recombinant DNA engineered in a T-region of a Ti plasmid. The vir region is located in a different plasmid within Agrobacterium.

Specific Patent Information

Patent Number Title, Independent Claims and Summary of Claims Assignee
US 5501967

  • Earliest priority – 26 July 1989
  • Filed – 6 July 1993
  • Granted – 26 March 1996
  • Expected expiry – 5 July 2013
 Title – Process for the site-directed integration of DNA into the genome of plants

Claim 1
A process for introducing a defined DNA sequence in a selected target locus of a nuclear plant cell genome by integrating at least a part of a recombinant DNA into said genome through homologous recombination at the target locus, comprising the steps of:a) coincubating plant protoplasts or plant cells under transforming conditions, with a strain of the genus Agrobacterium capable of T-region transfer and containing a plant transformation vector which comprises recombinant DNA of the formula:

wherein boxes 1 and 7 are T-DNA border sequences, and wherein one of box 1 or 7 may be absent; and wherein box 3 comprises a DNA sequence sufficiently homologous to a DNA sequence inside the target locus and sufficiently long to promote homologous recombination; and wherein box 4 comprises a DNA sequence not homologous to sequences occurring in the target locus; and wherein the lines connecting the boxes shown may comprise any number of nucleotides or basepairs, and
b) identifying protoplasts or cells that have obtained the said defined DNA sequence in their genomes at said target locus.
Claim 4
A process for introducing a defined mutation in a selected target locus of a nuclear plant cell genome by integrating at least a part of a recombinant DNA into said genome through homologous recombination at the target locus, comprising the steps of:a) coincubating plant protoplasts or plant cells under transforming conditions, with a strain of the genus Agrobacterium capable of T-region transfer and containing a plant transformation vector which comprises recombinant DNA of the formula:

wherein boxes 1 and 7 are T-DNA border sequences, and wherein one of box 1 or 7 may be absent; and wherein box 3 comprises a DNA sequence sufficiently homologous to a DNA sequence inside the target locus and sufficiently long to promote homologous recombination; and wherein box 4 comprises a DNA sequence sufficiently homologous to promote homologous recombination with a corresponding sequence in the target locus and wherein box (4) comprises the said defined mutation; and wherein the lines connecting the boxes shown may comprise any number of nucleotides or basepairs, and
b) identifying protoplasts or cells that have obtained the said defined mutation in their genome at the said target locus.
Claim 9
A recombinant DNA capable of introducing a defined mutation in a selected target locus of a nuclear plant cell genome through homologous recombination at the target locus, which recombinant DNA has the formula:

wherein box i comprises the said defined mutation; and wherein boxes 1 and 7 are T-DNA border sequences, and wherein one of box 1 or 7 may be absent; and wherein each of boxes 3 and 4 individually is a DNA sequence sufficiently homologous to a DNA sequence inside the target locus and sufficiently long to promote homologous recombination; and wherein the DNA sequences of the boxes 3 and 4 have the same 5′-to-3′ orientation, but wherein said boxes 3 and 4 are rearranged with respect to the homologous sequences in the target locus.

The patent US 5 501 967 claims

  • methods to introduce a defined sequence or a desired mutation into a target locus in a plant genome using constructs having at least one T-border, a sequence homologous to a sequence in the plant target locus, a sequence which is not homologous to the plant target locus sequence or which is homologous but which has a defined mutation;
  • a recombinant DNA having the defined mutation or sequence in a plant transformation vector within Agrobacterium . The recombinant DNA also contains:
    (i) at least one T-border, and
    (ii) a sequence homologous to a sequence in the plant target locus.
    The sequence to be inserted in the target locus is located between the homologous sequences in the vector. These sequences are in the same orientation but they are rearranged with respect to the orientation of the homologous sequence in the target locus.

Syngenta Mogen B.V.
EP 436007

  • Earliest priority – 26 July 1989
  • Filed – 26 July 1990
  • Granted – 26 March 1997
  • Expected expiry – 25 July 2010
 Title – Process for the site-directed integration of DNA into the genome of plants

Claim 1

A method for introducing a defined mutation in a selected target locus of a nuclear plant genome by integrating at least a part of a recombinant DNA into said genome through homologous recombination at the target locus, which recombinant DNA has the following general structure,

in which the boxes 1 and 7 and the connecting lines represent DNA sequences that are capable of functioning as a T-DNA border in the DNA transfer process, in which box 1 or 7 may be absent but not both, in which box 3 comprises a DNA sequence which is sufficiently homologous to a DNA sequence inside the target locus and sufficiently long to promote homologous recombination, in which box 4 represents a DNA sequence that is not homologous to sequences occurring in the target locus, and in which the lines connecting the boxes may represent any number of basepairs, or no basepairs, and identifying transformants having the defined mutation in said target locus using known methods.
Claim 8

A recombinant DNA capable of integrating a part of itself into the genome of a plant host via homologous recombination at the target locus,

in which the boxes 1 and 7 represent DNA sequences that are capable of functioning as a T-DNA border in the DNA transfer process, in which box 1 or 7 may be absent but not both, in which both box 3 and 4 represent DNA sequences which are sufficiently homologous and sufficiently long to promote homologous recombination with the target locus, in which the DNA sequences within the boxes have the same 5′ to 3′ orientation as in the target locus, but the order of the boxes themselves have been changed with respect to the situation in the target locus, resulting in insertion of box i in the target locus after homologous recombination, in which the lines connecting the boxes may represent any number of basepairs, or no basepairs.

Designated contracting States at the time of grant are: Austria, Belgium, Switzerland, Germany, Denmark, Spain, France, United Kingdom, Italy, Liechtenstein, Luxembourg, Netherlands, Sweden.
Remarks National phase entry of WO 1991/02070 in Japan (JP 3412134) has been published as granted on 3 June 2003.

Note: Patent information on this page was last updated on 24 February 2006.