Compositions and methods for locating preferred integration...

Details Compositions and methods for locating preferred integration... Compositions and methods for locating preferred integration...

1999-11-12

2003-04-22

Fox, David T. (Department: 1638)

Multicellular living organisms and unmodified parts thereof and

Method of introducing a polynucleotide molecule into or...

C800S288000, C800S320100, C435S468000, C435S419000, C435S462000

Reexamination Certificate

active

06552248

ABSTRACT:

FIELD OF THE INVENTION
The invention relates to the genetic modification of plants. Particularly, the control of gene integration and expression in plants is provided.
BACKGROUND OF THE INVENTION
Genetic modification techniques enable one to insert exogenous nucleotide sequences into an organism's genome. A number of methods have been described for the genetic modification of plants. All of these methods are based on introducing a foreign DNA into the plant cell, isolation of those cells containing the foreign DNA integrated into the genome, followed by subsequent regeneration of a whole plant. Unfortunately, such methods produce transformed cells that contain the introduced foreign DNA inserted randomly throughout the genome and often in multiple copies.
The random insertion of introduced DNA into the genome of host cells can be lethal if the foreign DNA happens to insert into, and thus mutate, a critically important native gene. In addition, even if a random insertion event does not impair the functioning of a host cell gene, the expression of an inserted foreign gene may be influenced by “position effects” caused by the surrounding genomic DNA. In some cases, the gene is inserted into sites where the position effects are strong enough to prevent the synthesis of an effective amount of product from the introduced gene. In other instances, overproduction of the gene product has deleterious effects on the cell.
Transgene expression is typically governed by the sequences, including promoters and enhancers, which are physically linked to the transgene. Currently, it is not possible to precisely modify the structure of transgenes once they have been introduced into plant cells. In many applications of transgene technology, it would be desirable to introduce the transgene in one form, and then be able to modify the transgene in a defined manner. By this means, transgenes could be activated or inactivated where the sequences that control transgene expression can be altered by either removing sequences present in the original transgene or by inserting additional sequences into the transgene.
For higher eukaryotes, homologous recombination is an essential event participating in processes like DNA repair and chromatid exchange during mitosis and meiosis. Recombination depends on two highly homologous extended sequences and several auxiliary proteins. Strand separation can occur at any point between the regions of homology, although particular sequences may influence efficiency. These processes can be exploited for a targeted integration of transgenes into the genome of certain cell types.
Even with the advances in genetic modification of higher plants, the major problems associated with the conventional gene transformation techniques have remained essentially unresolved as to the problems discussed above relating to variable expression levels due to chromosomal position effects and copy number variation of transferred genes. For these reasons, efficient methods are needed for targeting and control of insertion of nucleotide sequences to be integrated into a plant genome.
SUMMARY OF THE INVENTION
Compositions and methods for the targeted integration of nucleotide sequences into a transformed plant are provided. The compositions comprise transfer cassettes which are flanked by non-identical recombination sites.
The methods find use in targeting the integration of nucleotide sequences of interest to a specific chromosomal site, finding optimal integration sites in a plant genome, comparing promoter activity in transformed plants, engineering chromosomal rearrangements, and other genetic manipulation of plants.
Novel minimal recombination sites (FRT) are provided for use in the methods of the invention. Also provided are targeting cassettes and transgenic plants and plant cells containing corresponding non-identical recombination sites.


REFERENCES:
patent: 5677177 (1997-10-01), Wahl et al.
patent: 6010884 (2000-01-01), Griffiths et al.
patent: 6171861 (2001-01-01), Hartley et al.
patent: 6262341 (2001-07-01), Baszczynski et al.
patent: 6300545 (2001-10-01), Baszczynski et al.
patent: 6331661 (2001-12-01), Baszczynski et al.
patent: WO 92/15694 (1992-09-01), None
patent: WO 93/01283 (1993-01-01), None
patent: WO 93/17116 (1993-09-01), None
patent: WO 94/17176 (1994-08-01), None
patent: WO 95/00555 (1995-01-01), None
patent: WO 96/04393 (1996-02-01), None
patent: WO 97/09436 (1997-03-01), None
patent: WO 97/09439 (1997-03-01), None
patent: WO 97/13401 (1997-04-01), None
patent: WO 97/37012 (1997-10-01), None
patent: WO 97/47758 (1997-12-01), None
patent: WO 99/23202 (1999-05-01), None
patent: WO 99/55851 (1999-11-01), None
Lyznik, L. A. et al., “Activity of yeast FLP recombinase in maize and rice protoplasts.” 1993, Nucleic Acids Research, vol. 21, pp. 969-975.*
Albert, H. et al., “Site-specific integration of DNA into wild-type and mutant lox sites placed in the plant genome.” 1995, The Plant Journal, vol. 7, pp. 649-659.*
Lloyd, A.M., and R.W. Davis, “Functional Expression of the Yeast FLP/FRT Site-Specific Recombination System inNicotiana tobacum,” Mol. Gen Genet, 1994, pp. 653-657, vol. 242.
McLeod et al. Identification of the Crossover Site During FLP-Mediated Recombination in theSaccharomyces cerevisiaePlasmid 2&mgr;m Circle,Mol. Cell. Biol., Oct. 1986, pp 3357-3367, vol. 6(10), American Society for Microbiology, Cold Spring Harbor, New York.
Umlauf et al.The Functional Significance of DNA Sequence Structure in a Site-Specific Genetic Recombination Reaction, 1988, pp. 1845-1852, IRL Press Limited, Oxford, England.
Schlake et al. Use of Mutated FLP Recognition Target (FRT) Sites for the Exchange of Expression Cassettes at Defined Chromosomal Loci,Biochem., 1994, vol. 33(43), pp. 12746-12751, American Chemical Society.
Osborne, B., et al., “A System for Insertional Mutagenesis and Chromosomal Rearrangement Using the Ds Transposon and Cre-lox,”The Plant Journal, 1995, pp. 687-701, vol. 7(4).
Karreman et al. (1996) “On the Use of Double FLP Recognition Targets (FRTs) in the LTR of Retroviruses for the Construction of high Producer Cell Lines”,Nucleic Acids Research 24(9):1616-1624.
Storici et al. (1997) “Molecular Engineering with the FRT Sequence of the Yeast 2 &mgr;mPlasmid: [ciro] Segregant Enrichment By Counterselection for 2 &mgr;m Site-Specific Recombination”,Gene 195:245-255.
Czakó et al. (1997) “Negative Selection Markers for Plants”, Technology Transfer of Plant Biotechnology, Chapter 6, Edited by Peter M. Gresshoff, Plant Molecular Genetics, Institute of Agriculture, Center for Legume Research, The University of Tennessee, Knoxville, Tennessee, CRC Press, pp. 67-93.
Dasgupta et al. (1991) “Rice Tungro Bacilliform Virus DNA Independently Infects Rice After Agrobacterium-mediated Transfer”,Journal of General Virology72:1215-1221.
Grimsley et al. (1988) “Meristematic Tissues of Maize Plants are Most Susceptible to Agroinfection with Maize Streak Virus”,Bio/Technology 6:185-189.
Louie (1995) “Vascular Puncture of Maize Kernels for the Mechanical Transmission of Maize White Line Mosaic Virus and Other Viruses of Maize”,Phytopathology 85)2):139-143.
Scholthof et al. (1996) “Plant Virus Gene Vectors For Transient Expression Of Foreign Proteins In Plants”,Annu. Rev. of Phytopathol. 34:299-323.
Timmermans et al. (1992) “Trans Replication and High Copy Numbers of Wheat Dwarf Virus Vectors in Maize Cells”,Nucleic Acids Research 20(15):4047-4054.
Ugaki et al. (1991) “Replication of aGeminivirus Derived Shuttle Vector in Maize Endosperm Cells”,Nucleic Acids Research 19(2):371-377.
Snaith et al., Multiple Cloning Sites Carrying loxP and FRT Recognition Sites for the Cre and Flp Site-Specific Recombinases, Gene, 1995, pp. 173-174, vol. 166.
Ow et al., Genome Manipulation Through Site-Specific Recombination, Critical Reviews in Plant Sciences, (1995), pp 239-261, vol. 14(3).
Senecoff et al. Directionality in FLP Protein-Promoted Site-Specific Recombination Is Mediated by DNA-DNA Pairing,J. Biol. Chem., Jun. 5, 1986, pp. 7380-7386, vol. 261(16), The American Society of Biologic

Affiliated with

Also associated with

Rating

Compositions and methods for locating preferred integration... does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Compositions and methods for locating preferred integration..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Compositions and methods for locating preferred integration... will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-3066735