Electroporation-mediated molecular transfer in intact plants
AssigneeGenetronics, Inc.
InventorsS. B. Dev, Yasuhiko Hayakawa
Life of this patent
Estimating the term…
Abstract
A method for producing a genetically modified plant by introducing a polynucleotide to an intact plant or plant cell(s) via electroporation, in the absence of cell wall-degrading enzymes. Genetically engineered plants produced by the method of the invention are also provided. The invention also provides a method for producing a polypeptide in an intact plant cell, including plant tissue or a whole plant by introducing a biologically active polypeptide directly into the plant.
Claims30
A method of producing a genetically modified plant, the method comprising:
- contacting intact plant tissue with a polyucleotide wherein the polynucleotide is operably associated with a promoter;
- applying a pulsed electrical field via electroporation to the plant tissue, under conditions and for sufficient time to allow uptake of the polynucleotide, wherein the length of the electrical impulse applied is from about 1.0 .mu.sec to 20 msec and wherein the plant tissue is not pretreated with a cell membrane-penetrating agent or a lipophilic agent;
- and
- expressing the polynucleotide in cell(s) of the tissue.
A method for introducing a heterologous polypeptide into intact plant tissue comprising:
- contacting the plant tissue with the polypeptide to be introduced, wherein the introduced polypeptide is biologically active;
- applying a pulsed electrical field via electroporation to the plant tissue, under conditions and for sufficient time to allow uptake of the polypeptide into the cells of the tissue;
- and
- recovering the polypeptide from the plant tissue.
A method of modulating gene expression in intact plant tissue, comprising:
- contacting plant tissue with a modulatory amount of polynucleotide;
- applying a pulsed electrical field via electroporation to the plant tissue, under conditions and for sufficient time to allow uptake of the polynucleotide;
- and
- modulating gene expression in the plant tissue.
Dependencies are inferred from the claim text.