Rutgers, The State University of New Jersey

University of Minnesota

University of Medicine and Dentistry of New Jersey

Nilgun Ereken Tumer

Rong Di

Katalin A Hudak

Fatih M Uckun

Jonathan D Dinman

Thomas Leustek

Xiao-Ping Li

Mohamed Anwar Bin-Umer

John Edward McLaughlin

Bijal Parikh

Compositions and methods are disclosed for modulating disease resistance in plants.

Plant genes that confer resistance to trichothecene mycotoxins and fusarium head blight

Compositions and methods for treatment of toxin poisoning are disclosed.

Ricin ribosome binding protein compositions and methods of use thereof

A method of treating or inhibiting hepatitic C virus (HCV). The method comprises administering an effective amount of at least one pokeweed antiviral protein (PAP) mutant alone or in combination with other anti-HCV agents.

Method of treating hepatitis C virus

Disclosed are specific mutants of L3 and transgenic plants that produce them. The plants exhibit increased resistance to fungal toxins that target ribosomal L3 protein. Also disclosed are transgenic plants that co-produce L3 mutant and an RIP protein, and exhibit increased resistance to various fungal toxins and viruses, while reducing toxicity normally associated with production of the RIP. Uses of the L3 mutants in animals are further disclosed.

Transgenic plants expressing L3 delta proteins are resistant to trichothecene fungal toxins

Disclosed are PAP mutants that are less toxic than wild type PAP and that exhibit depurination activity. Also disclosed are transgenic plants that procedure the PAP mutants, and methods for preparing the plants. Further disclosed are bioconjugates containing the PAP mutants, pharmaceutical compositions containing the bioconjugates, and methods of administering the compositions to treat disease.

C-terminally truncated non-cytotoxic PAP mutants

Disclosed are transgenic plants containing an exogenous nucleic acid encoding an L3 protein. The plant exhibits increased resistance to viruses and/or fungi that infect plants. The L3 proteins include wild-type proteins, spontaneously occurring mutants and non-naturally occurring L3 mutants. Also disclosed are methods of reducing the toxicity of single-chain ribosome inhibitory proteins in cells, e.g., yeast, plant and animal cells, by co-administering the L3 protein with the RIP. Further disclosed are non-naturally occurring L3 mutants that (a) substantially fail to bind single-chain RIPs that bind endogenous L3 proteins, (b) are unable to maintain M1 killer virus, (c) promote altered programmed ribosomal frameshift efficiency, (d) exhibit resistance to peptidyltransferase inhibitors, and combinations of any of (a)&#x2013;(d).

Virus-resistant transgenic plants expressing L3

Disclosed are transgenic plants having edible portions that produce methional during processing. The plants contain increased methionine levels such that upon processing of the edible portion(s), methional levels are increased and lead to food products that possess improved flavor stability and/or quality. Plants of the Solanaceous family e.g., potato, tomato and eggplant, and other methional-producing plants including maize and soybean, are preferred plants. Several ways of genetically engineering plants to produce increased free Met levels are disclosed, with introduction of a non-native nucleic acid encoding cystathionine gamma synthase (CGS) and tissue-specific expression of an anti-sense S-adenosyl-methionine synthetase being preferred. Also disclosed are methods for selecting transformed plant cells using ethionine and CGS as the selection agent and marker gene respectively.

Method for selecting transformed plant cells using ethionine and cystathionine gamma synthase as the selection agent and marker gene

Disclosed are PAP mutants having reduced phytotoxicity compared to wild-type PAP, and which confer broad spectrum resistance to viruses and/or fungi in plants. One group of PAP mutants is characterized by at least one amino acid substitution in the N-terminus of mature PAP, such as the Glycine 75 residue or the Glutamic acid 97 residue; two groups of additional PAP mutants are characterized by truncations in the N-terminal region of mature PAP and truncations or amino acid substitutions in the C-terminal region of mature PAP, respectively; and a further group are enzymatically inactive which still exhibit anti-fungal properties. Also disclosed are DNA molecules encoding the PAP mutants, mutant PAP DNA constructs, and transgenic seed and plants containing the DNAs. Further disclosed are methods for identifying PAP mutants having reduced phytotoxicity, as well as isolated and purified PAP mutants identified by the method.

Pap mutants that exhibit anti-viral and/or anti-fungal activity in plants

Biotherapeutic agents are provided which comprise recombinant PAP or a biologically equivalent variant or mutant thereof, linked to a targeting moiety which are effective for the treatment of certain human diseases. The invention further provides a process for producing the biotherapeutic agents as well as a method which utilizes the disclosed biotherapeutic agents to systemically treat cancer patients.

Biotherapeutic agents comprising recombinant PAP and PAP mutants

Disclosed are PAP mutants having reduced phytotoxicity compared to wild-type PAP, and which retain wild-type PAP biological activity in plants. One group of PAP mutants is characterized by at least one amino acid substitution in the N-terminus of mature PAP, such as the Glycine 75 residue or the Glutamic acid 97 residue. Another group of preferred PAP mutants is characterized by mutations such as truncations in the C-terminal region of mature PAP. PAP mutants having from at least about 26 to about 76 mature PAP amino acids (not counting the 29-amino acid C-terminal extension of wild-type PAP) exhibit reduced phytotoxicity and retain PAP biological activity in plants. The disclosed PAP mutants may include the 22-amino acid N-terminal signal sequence and/or the C-terminal extension of wild-type PAP. Also disclosed are DNA molecules encoding the PAP mutants. The DNAs can be operably linked to a promoter functional in given host cells such as plants, and stably transformed into a vector functional in said cells. Procaryotic or eucaryotic hosts, e.g., yeast or plants, stably transformed with a mutant PAP-encoding DNA are further disclosed, as well as protoplasts stably transformed with the DNAs. Transgenic plants and seed containing the DNAs are also provided. The transgenic plants exhibit broad spectrum virus resistance. They include monocots, such as cereal crops, and dicot plants. Further disclosed is a method for identifying a PAP mutant having reduced phytotoxicity and which retains PAP biological activity. The method involves the steps of providing a eucaryotic cell stably transformed with a mutagenized PAP-encoding DNA molecule, wherein the DNA molecule is operably linked to an inducible promoter functional in eucaryotic cells. The thus-transformed cell is cultured in a suitable medium, and after a predetermined time, an inducer is added to the medium to cause expression of the DNA molecule. A determination is then made as to whether any cultured cells survive the induction of expression of the DNA molecule. The presence of which indicates the presence of a PAP mutant having reduced phytotoxicity so that the biological activity of the PAP mutant encoded by the mutagenized DNA can then be determined. Any PAP mutants which also exhibit broad spectrum virus resistance in an in vivo or in vitro assay would be considered as PAP mutants which retain PAP biological activity in plants. Isolated and purified PAP mutants identified by the aforesaid process are also provided.

Growing community of inventors

Belle Mead, NJ, United States of America

Nilgun Ereken Tumer