CA2200496A1 - Genetic transformation using a parp inhibitor - Google Patents

Genetic transformation using a parp inhibitor

Info

Publication number
CA2200496A1
CA2200496A1 CA002200496A CA2200496A CA2200496A1 CA 2200496 A1 CA2200496 A1 CA 2200496A1 CA 002200496 A CA002200496 A CA 002200496A CA 2200496 A CA2200496 A CA 2200496A CA 2200496 A1 CA2200496 A1 CA 2200496A1
Authority
CA
Canada
Prior art keywords
cells
plant
cell
foreign dna
gene
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CA002200496A
Other languages
French (fr)
Other versions
CA2200496C (en
Inventor
Marc De Block
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Bayer CropScience NV
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from EP95401844A external-priority patent/EP0757102A1/en
Application filed by Individual filed Critical Individual
Publication of CA2200496A1 publication Critical patent/CA2200496A1/en
Application granted granted Critical
Publication of CA2200496C publication Critical patent/CA2200496C/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8287—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for fertility modification, e.g. apomixis
    • C12N15/8289—Male sterility

Landscapes

  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Zoology (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • Plant Pathology (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Cell Biology (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention concerns a process for producing transgenic eucaryotic cells, particularly plants, which comprises: contacting a culture of untransformed cells with an inhibitor of poly-(ADP-ribose) polymerase for a period of time sufficient to reduce the response of the cultured cells to stress and to reduce their metabolism. The untransformed cells are then contacted with foreign DNA comprising at least one gene of interest under conditions in which the foreign DNA is taken up by the untransformed cells and the gene of interest is stably integrated in the nuclear genome of the untransformed cells to produce the transgenic cells. Optionally, the transgenic cells are recovered from the culture. Preferably, the inhibitor is niacinamide, preferably at a concentration of about 200 mg/l to 500 mg/l and the untransformed cells are cultured in a medium containing the inhibitor for a period of time of approximately 3 to 14 days prior to the contacting with the foreign DNA. The invention also relates to a plant having in the nuclear genome of its cells foreign DNA integrated only in the regions of the nuclear genome that are transcriptionally active in cells of the plant when the cells are treated with an effective amount of a PARP inhibitor for a period of time sufficient to reduce cell metabolism to a state where gene expression is essentially limited to genes expressed irrespective of the differentiated or physiological condition of the cell.

Claims (23)

1. A process for producing transgenic eucaryote cells which comprises:
contacting a culture of untransformed cells with an inhibitor of poly-(ADP-ribose) polymerase, prior to transformation, for a period of time sufficient to reduce the response of the cultured cells to stress and to reduce the metabolism of said cultured cells, particularly to reduce the electron flow in the mitochondrial electron transport chain; contacting said transformed cells with foreign DNA comprising at least one gene of interest under conditions in which said foreign DNA is taken up by said untransformed cells and said gene of interest is stably integrated in the nuclear genome of said untransformed cells to produce said transgenic cells; and optionally recovering said transgenic cells from said culture.
2. The proces of claim 1, wherein said eucaryotic cells are plant cells.
3. The process of claim 1 or 2, wherein said inhibitor is niacinamide, preferably at a concentration of about 150 mg/l to 1000 mg/l, more preferably at a concentration of about 200 mg/l to 500 mg/l, particularly at a concentration of about 250 mg/l.
4. The process of any one of claims 1 to 3, wherein said untransformed cells are cultured in a medium containing said inhibitor for a period of time of approximately 2 to 28 days, preferably approximately 3 to 14 days, particularly approximately 4 days prior to the contacting with said foreign DNA.
5. The process of any one of claims 1 to 4, wherein said cells contacted with said foreign DNA are further cultured in a medium containing said inhibitor for a period of time of approximately 1 to 14 days preferably 2 to 4 days after contacting with said foreign DNA.
6. A process for increasing the frequency of obtaining transgenic plant cells which comprises:
contacting untransformed plant cells with foreign DNA comprising at least one gene of interest under conditions in which said foreign DNA is taken up by said untransformed cells and said gene of interest is stably integrated in the nuclear genome of said untransformed cells to produce said transgenic cells contacting cells with an inhibitor of poly-(ADP-ribose); and further culturing said cells in a medium containing said inhibitor for a period of time of approximately 1 to 14 days preferably 1 to 4 days, particularly 1 day after contacting with said foreign DNA.
7. The proces of any of claims 1 to 6, wherein said gene of interest comprises a promoter that directs expression selectively in certain cells or tissues of an eucaryotic organism.
8. The process of any one of claims 2 to 7, wherein said gene of interest comprises a promoter that directs expression selectively in stamen cells, particularly anther cells of a plant.
9. The process of claim 7 or 8, wherein said gene of interest encodes a protein that when produced in a cell of an eucaryotic organism, kills or disables said cell.
10. The process of claim 9, wherein said gene of interest encodes a ribonuclease particularly barnase.
11. The process of any one of claims 1 to 10, wherein a transgenic organism having said foreign DNA with said at least one gene of interest stably integrated in its genome is obtained from said transformed eucaryotic cell.
12. The proces of claim 11 wherein said organism is a plant which is obtained by regeneration from a transformed plant cell.
13. The transgenic organism obtained by the process of claim 11 or 12.
14. A plant having foreign DNA integrated in the nuclear DNA of its cells only in the regions of said nuclear DNA that are transcriptionally active in said cells of said plant when said cells are treated with an effective amount of a PARP inhibitor for a period of time sufficient to reduce cell metabolism to a state where gene expression is essentially limited to genes expressed irrespective of the differentiated or physiological condition of the cell.
15. The plant according to claim 14, wherein said integration of the foreign DNA in said transcriptionally active region is verified by measuring the level of expressed mRNA corresponding to this foreign DNA when said cells are incubated in a medium containing a PARP-inhibitor.
16. The plant according to claim 14, wherein said transcriptionally active regions of the genome of said plant include regions which are minimally affected by cell differention or cell physiological and biochemical changes caused by external factors such as environmantal conditions especially stress conditions.
17. The plant or plant cell according to any one of the preceeding claims, wherein said plant or plant cell is a monocotyledonous plant or plant cell.
18. The plant or plant cell according to claim 17, wherein said plant or plant cell is a cereal plant or plant cell.
19. The plant of plant cell acccording to claim 18, wherein said plant or plant cell is wheat or a wheat cell.
20. The plant according to any one of the preceeding claims, wherein said foreign DNA comprises a DNA sequence expressed selectively in specific tissues of said plant.
21. The plant of claim 20, wherein said foreign DNA comprises a DNA
sequence encoding a cytotoxic molecule.
22. The plant of claim 21, wherein said foreign DNA comprises a DNA
sequence encoding barnase.
23. A eucaryotic cell having foreign DNA integrated in its nuclear DNA only in the regions of said nuclear DNA that are transcriptionally active in said cell when said cell is treated with an effective amount of a PARP inhibitor for a period of time sufficient to reduce cell metabolism to a state where gene expression is essentially limited to genes expressed irrespective of the differentiated or physiological condilion of the cell.
CA2200496A 1995-08-04 1996-07-31 Genetic transformation using a parp inhibitor Expired - Fee Related CA2200496C (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
GB95401844.6 1995-08-04
EP95401844A EP0757102A1 (en) 1995-08-04 1995-08-04 Genetic transformation using a PARP inhibitor
PCT/EP1996/003366 WO1997006267A2 (en) 1995-08-04 1996-07-31 Genetic transformation using a parp inhibitor

Publications (2)

Publication Number Publication Date
CA2200496A1 true CA2200496A1 (en) 1997-02-20
CA2200496C CA2200496C (en) 2010-04-20

Family

ID=42115799

Family Applications (1)

Application Number Title Priority Date Filing Date
CA2200496A Expired - Fee Related CA2200496C (en) 1995-08-04 1996-07-31 Genetic transformation using a parp inhibitor

Country Status (1)

Country Link
CA (1) CA2200496C (en)

Also Published As

Publication number Publication date
CA2200496C (en) 2010-04-20

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