Chemistry: molecular biology and microbiology – Process of mutation – cell fusion – or genetic modification – Introduction of a polynucleotide molecule into or...
Patent
1997-05-15
2000-06-13
Fox, David T.
Chemistry: molecular biology and microbiology
Process of mutation, cell fusion, or genetic modification
Introduction of a polynucleotide molecule into or...
435199, 435418, 435419, 435420, 435430, 435469, 800278, 800287, 800294, 800300, 800303, 800306, 800320, 8003203, C12N 1582, C12N 1584, C12N 1555, A01H 102
Patent
active
060748761
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
This invention is related to tissue culture of eucaryotic cells and improved techniques to obtain genetically transformed eucaryotic cells and organisms, such as transgenic plant cells or plants, by lowering the stress reaction of cultured eucaryotic cells prior to contacting the cells with foreign DNA, particularly by specific inhibition of poly-(ADP-ribose) polymerase.
BACKGROUND OF THE INVENTION
Over the years many techniques for the genetic transformation of higher organisms (animals and plants) have been developed. In these techniques it is the ultimate goal to obtain a transgenic organism, e.g. a plant, in which all cells contain a foreign DNA comprising a gene of interest (the so-called transgene) stably integrated in their genome, particularly their nuclear genome.
Transformation is a complex process which always involves the contacting of starting cells with a DNA, usually a DNA comprising foreign gene(s) of interest. The contacting of the cells with the DNA is carried out under conditions that promote the uptake of the DNA by the cells and the integration of the DNA, including the gene(s) of interest into the genome of the cell.
Starting cells for transformation are usually cells that have been cultured in vitro for some time. After contacting the cells with the DNA, the transformed cells generally need to be cultured in vitro for a certain period in order to separate the transformed cells from the non-transformed cells and, in the case of plants, to regenerate transformed plants from the transformed cells. Indeed, complete plants can be regenerated from individual transformed cells thus ensuring that all cells of the regenerated plant will contain the transgene.
In many plants, genetic transformation can be achieved by using the natural capacity of certain Agrobacterium strains to introduce a part of their Ti-plasmid, i.e. the T-DNA, into plant cells and to integrate this T-DNA into the nuclear genome of the cells. It was found that the part of the Ti-plasmid that is transferred and integrated is delineated by specific DNA sequences, the so-called left and right T-DNA border sequences and that the natural T-DNA sequences between these border sequences can be replaced by foreign DNA (European Patent Publication "EP" 116718; Deblaere et al, 1987 Meth.Enzymol. 153:277-293).
Certain plant species have proven to be recalcitrant to Aqrobacterium mediated transformation and in these species, as well as in animals, genetic transformation has been achieved by means of direct gene transfer by which DNA is inserted into the cells by physical and/or chemical means, such as by electroporation, by treatment of the cells with polyethyleneglycol (PEG), by bombardment of the cells with DNA-coated microprojectiles, etc. (WO 92/09696; Potrykus et al, 1991, Annu.Rev.Plant Physiol.Plant Mol.Biol. 42:205-225).
Genetic transformation of eucaryotic cells is generally a random event, i.e. the transgene is integrated in the genome at random positions. Often several copies (or parts of copies) of the transforming DNA are integrated in a single position, and/or at different positions, resulting in a transformed cell containing multiple copies of the transgene.
The expression of the transgene is known to be influenced by its position in the genome. For instance, a foreign DNA, when introduced in a plant cell appears to integrate randomly in the plant genome. Examination of independently transformed plants has shown a high degree of variability (up to 1 00-fold) in the expression level of the introduced gene. Several studies have shown no correlation between this "between-transformant variability" and the copy number of the introduced DNA at a given locus. It has been suggested that some of the variability in expression of introduced genes in transgenic plants is a consequence of "position effects" caused by influences of adjacent plant genomic DNA. Other factors that could contribute to the variability in expression are physiological variability of the plant material, differences in the number of
REFERENCES:
"Illegitimate and homologous recombination in mammalian cells: differential sensitivity to an inhibitor of poly(ADP-ribosylation)" by B. C. Waldman et al., Nucleic Acids Research, vol. 18, No. 20 (1990) pp. 5981-5988.
"Secondary Metabolism In Cultured Red Beet Beta-Vugaris L. Cells Differential Regulation of Betaxanthin and Betacyanin Biosynthesis" by P-A Girod et al, Biological Abstracts, vol. 92 (1991), Abstract No. 16692.
"Reduced Position Effect in Mature Transgenic Plants Conferred by the Chicken Lysozyme Matrix-Associated Region" by L. Mlynarova et al, The Plant Cell, vol. 6 (1994) pp. 417-426.
"Scaffold Attachment Regions Increase Reporter Gene Expression in Stably Transformed Plant Cells" by G. C. Allen et al, The Plant Cell, vol. 5 (1993) pp. 603-613.
"Transgene expression variability (position effect) of CAT and GUS reporter genes driven by linked divergent T-DNA promoters" by C. Peach et al, Plant Molecular Biology, vol. 17 (1991) pp. 49-60.
"In vivo random beta-glucuronidase gene fusions in Arabidopsis thaliana" by S. Kertbundit et al, Proceedings of the National Academy of Sciences of USA, vol. 88 (1991) pp. 5212-5216.
"Assessment of promoter trap as a tool to study zygotic embryogenesis in Arabidopsis thaliana" by M. Devic et al, Compes Rendus De L'Academie Des Sciences Serie III Sciences De La Vie. 318 (1) (1995), pp. 121-128.
"Identification of Arabidopsis thaliana sequences responsive to low temperature and abscisic acid by T-DNA tagging and in-vivo gene fusion" by A. Mandal et al, Chemical Abstracts, vol. 123, No. 21, (1995), Abstract No. 276968.
Waldman et al. Illegitimate and homologous recombination in mammalian cells: differential sensitivity to an inhibitor of poly (ADP-ribosylation). Nucleic Acids Research. 18(20):5981-5988, 1988.
Strain. Inhibitors of ADP-ribosyl transferase enhance the transformation of NIH3T3 cells following transfection with SV40 DNA. Exp. Cell Res. 159:531-535, Aug. 1985.
Fox David T.
Plant Genetic Systems N.V.
LandOfFree
Genetic transformation using a PARP inhibitor does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Genetic transformation using a PARP inhibitor, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Genetic transformation using a PARP inhibitor will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2067708