The patent's inventors are Samboju,
This patent was filed on
From the background information supplied by the inventors, news correspondents obtained the following quote: "Nanoparticles have unique properties that have been exploited to deliver DNA to specific animal cells. It has been found that when certain DNA-coated nanoparticles are incubated with cells not having a cell wall, the cells take up the nanoparticles and begin expressing genes encoded on the DNA. Semi-conductor nanoparticles (e.g., quantum dots ('QDs')) within the size range of 3-5 nm have also been used as carriers to deliver molecules into cells. DNA and proteins can be linked to certain ligands attached to the QD surface. See e.g., Patolsky, et al. (2003) J. Am. Chem. Soc. 125:13918. Carboxylic acid- or amine-coated QDs can be cross-linked to molecules containing a thiol group, see, e.g., Dubertret et al. (2002) Science 298:1759; Akerman, et al. (2002) Proc. Natl. Acad. Sci.
"Delivery of foreign nucleic acid molecules to plants is challenging due to the presence of plant cell walls. Current methods rely on invasive delivery for genetic transformation of plants. In plant cells, the cell wall is a barrier against the delivery of exogenously applied molecules. Many invasive cell delivery methods, for example, biolisitic delivery (gene gun), microinjection, electroporation, and Agrobacterium-mediated transformation, have been employed to achieve gene and small molecule delivery into walled plant cells, but delivery of proteins has only been achieved by microinjection. Where nanoparticle delivery of nucleic acid molecules to plant cells is desired, the cell wall is removed before the addition of the particles to protoplasts of plant. See, for example, Torney, et al. (2007) Nature Nanotechnol. 2:295-300."
Supplementing the background information on this patent, NewsRx reporters also obtained the inventors' summary information for this patent: "Described herein are methods and compositions for use of nanoparticles and linearized nucleic acid molecules for introducing a molecule of interest into a plant cell having a cell wall. Some embodiments of methods of the disclosure may be used to produce a stably-transformed genetically modified fertile plant. In some embodiments, the distinctive properties of linear nucleic acid molecules allow the delivery of specific gene sequences of interest without extraneous nucleic acid sequences that may have regulatory consequences for a transgenic target organism.
"In some embodiments, nanoparticles may be PEGylated with linear nucleic acid molecules. In particular embodiments, the nanoparticles may be semi-conductor nanoparticles, such as quantum dots ('QDs'). In some embodiments, the linear nucleic acid molecules may be linearized plasmid DNA. In other embodiments, the linear nucleic acid molecules may comprise sequences encoding Phosphoinothricin-N-acetyltransferase (PAT) and/or Yellow fluorescence protein (YFP).
"Also disclosed are methods for introducing a molecule of interest into a plant cell having a cell wall, wherein the methods may comprise providing the plant cell having a cell wall; coating the surface of nanoparticles with PEG to produce 'PEGylated' nanoparticles; coating the PEGylated nanoparticles with at least one linear nucleic acid molecule of interest; placing the plant cell having a cell wall and the PEGylated nanoparticles coated with the linear nucleic acid molecule(s) of interest in contact with each other; and allowing uptake of the nanoparticle and the linear nucleic acid molecule(s) of interest into the cell comprising a cell wall.
"Further disclosed are methods for introgressing a trait into a plant. In some embodiments, the method may comprise providing a plant cell; coating the surface of nanoparticles with PEG to produce PEGylated nanoparticles; coating the PEGylated nanoparticles with a means for expressing the trait in the plant; placing the plant cell and the PEGylated nanoparticles coated with means for expressing the trait in the plant in contact with each other; allowing uptake of the nanoparticle and the means for expressing the trait in the plant into the plant cell to produce a transformed plant cell; regenerating a whole plant from the transformed plant cell; and propagating the plant. In some embodiments, a trait that may be introgressed according to methods of the invention include a trait selected from, without limitation: male sterility; herbicide resistance; insect resistance; and resistance to bacterial disease, fungal disease, and/or viral disease.
"Also disclosed are methods of the invention may be used for in planta transformation of a plant. In some embodiments, the plant may be selected from plants of the genus, Arabidopsis, for example, A. thaliana. In particular embodiments, a plant transformed by in planta transformation may be selected from A. thaliana plants of the
"Additionally disclosed are genetically modified (GM) plant cells and methods for generating them, wherein the plant cells have one or more nucleic acids introduced therein via methods of the present invention. In some embodiments, a plasmid comprising at least one gene of interest and a selectable marker may be in introduced into a plant cell having a cell well via a nanoparticle according to the present invention. In further embodiments, stable transformants may be selected that have stably integrated at least one gene of interest and/or the selectable marker. In alternative embodiments, a plant cell now comprising at least one gene of interest may be propagated to produce other cells comprising a molecule of interest. In other embodiments, plant cells now comprising a molecule of interest may be a regenerable cell that may be used to regenerate a whole plant including the molecule of interest.
"Further disclosed are methods of creating regenerable plant cells comprising a molecule of interest for use in tissue culture. The tissue culture may be capable of regenerating plants having substantially the same genotype as the regenerable cells. The regenerable cells in such tissue cultures may be, for example, embryos; protoplasts; meristematic cells; callus; pollen; leaves; anthers; roots; root tips; flowers; seeds; pods; or stems. Still further, some embodiments provide plants regenerated from the tissue cultures of the invention.
"Further disclosed are methods for generating stabilized plant lines comprising a desired trait or nucleic acid molecule of interest, wherein the desired trait or nucleic acid molecule of interest may be first introduced by uptake of a nanoparticle across a plant cell wall. Methods of generating stabilized plant lines are well known to one of ordinary skill in the art, and may include techniques such as, but not limited to, selfing; backcrossing; hybrid production; crosses to populations; and the like. Thus, also disclosed are plants and plant cells comprising a desired trait or nucleic acid molecule of interest first introduced into the plant cell (or its predecessors) by uptake of a nanoparticle across a cell wall. Plant cells comprising a desired trait or nucleic acid molecule of interest first introduced into the plant or cell (or its predecessors) by uptake of a nanoparticle across a cell wall can be used in crosses with other, different, plant cells to produce first generation (F.sub.1) hybrid cells; seeds; and/or plants with desired characteristics.
"In addition to the exemplary aspects and embodiments described above, further aspects and embodiments will become apparent in view of the following descriptions."
For the URL and additional information on this patent, see: Samboju,
Keywords for this news article include: Peptides, Proteins, Cell Wall, Agriculture, Amino Acids,
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