Multiple Inducible Gene Regulation System

1 .- 20 . (canceled)

21 . A non-human organism comprising a multiple inducible gene modulation system comprising two individually operable heterologous gene modulation systems,

wherein each individually operable heterologous gene modulation system comprises:

a) a first gene expression cassette comprising a polynucleotide sequence that encodes a polypeptide comprising:

A) a DNA binding domain that recognizes a response element associated with a gene of interest;

B) an ecdysone receptor ligand binding domain; and

b) a second gene expression cassette comprising a polynucleotide sequence comprising a promoter operably linked to a polynucleotide sequence that encodes a second polypeptide comprising a nuclear receptor ligand binding domain and a transactivation domain; and

wherein each individually operable heterologous gene modulation system is orthogonal to the other individually operable heterologous gene modulation system present in the multiple inducible gene modulation system.

22 . The non-human organism of claim 21 , wherein said organism is a transgenic organism.

23 . The non-human organism of claim 21 , wherein said organism is an isolated cell.

24 . The non-human organism of claim 21 , wherein said nuclear receptor ligand binding domain is selected from the group consisting of a vertebrate retinoid X receptor ligand binding domain; an invertebrate retinoid X receptor ligand binding domain; an ultraspiracle protein ligand binding domain; and a chimeric ligand binding domain comprising two polypeptide fragments, wherein the first polypeptide fragment is from a vertebrate retinoid X receptor ligand binding domain, an invertebrate retinoid X receptor ligand binding domain, or an ultraspiracle protein ligand binding domain, and the second polypeptide fragment is from a different vertebrate retinoid X receptor ligand binding domain, invertebrate retinoid X receptor ligand binding domain, or an ultraspiracle protein ligand binding domain.

25 . The non-human organism of claim 21 , wherein said ecdysone receptor ligand binding domain is selected from the group consisting of a Lepidopteran ecdysone receptor ligand binding domain, a Dipteran ecdysone receptor ligand binding domain, an Arthropod ecdysone receptor ligand binding domain, an Orthopteran ecdysone receptor ligand binding domain, a Homopteran ecdysone receptor ligand binding domain, a Hemipteran ecdysone receptor ligand binding domain, a spruce budworm Choristoneura fumiferana ecdysone receptor ligand binding domain, a yellow meal worm Tenebrio molitor ecdysone receptor ligand binding domain, a tobacco hornworm Manduca sexta ecdysone receptor ligand binding domain, a tobacco budworm Heliothies virescens ecdysone receptor ligand binding domain, a golmidge Chironomus tentans ecdysone receptor ligand binding domain, a silkworm Bombyx mori ecdysone receptor ligand binding domain, a squinting bush brown Bicyclus anynana ecdysone receptor ligand binding domain, a buckeye Junonia coenia ecdysone receptor ligand binding domain, a fruit fly Drosophila melanogaster ecdysone receptor ligand binding domain, a yellow fever mosquito Aedes aegypti ecdysone receptor ligand binding domain, a blowfly Lucilia capitata ecdysone receptor ligand binding domain, a sheep blowfly Lucilia cuprina ecdysone receptor ligand binding domain, a blowfly Calliphora vicinia ecdysone receptor ligand binding domain, a Mediterranean fruit fly Ceratitis capitata ecdysone receptor ligand binding domain, a locust Locusta migratoria ecdysone receptor ligand binding domain, an aphid Myzus persicae ecdysone receptor ligand binding domain, a fiddler crab Celuca pugilator ecdysone receptor ligand binding domain, an ixodid tick Amblyomma americanum ecdysone receptor ligand binding domain, and a white fly Bamecia argentifoli ecdysone receptor ligand binding domain.

26 . The non-human organism of claim 25 , wherein said ecdysone receptor ligand binding domain is a spruce budworm Choristoneura fumiferana ecdysone receptor ligand binding domain.

27 . The non-human organism of claim 21 , wherein said DNA binding domain is selected from the group consisting of a GAL4 DNA binding domain, a LexA DNA binding domain, a transcription factor DNA binding domain, a Group H nuclear receptor member DNA binding domain, a steroid/thyroid hormone nuclear receptor superfamily member DNA binding domain, a bacterial LacZ DNA binding domain, and an ecdysone receptor DNA binding domain.

28 . The non-human organism of claim 21 , wherein said transactivation domain is selected from the group consisting of a Group H nuclear receptor member transactivation domain, a steroid/thyroid hormone nuclear receptor transactivation domain, a polyglutamine transactivation domain, a basic or acidic amino acid transactivation domain, a VP16 transactivation domain, a GAL4 transactivation domain, an NF-KB transactivation domain, a BP64 transactivation domain, a B42 acidic transactivation domain, and a p65 transactivation domain.

29 . The non-human organism of claim 21 , wherein said organism is a prokaryote.

30 . The non-human organism of claim 21 , wherein said organism is an invertebrate.

31 . The non-human organism of claim 21 wherein said organism is selected from the group consisting of a bacterium, a fungus and a yeast.

32 . The non-human organism of claim 31 , wherein said bacterium is selected from the group consisting of Synechocystis, Synechococcus, Salmonella, Bacillus, Acinetobacter, Rhodococcus, Streptomyces, Escherichia, Pseudomonas, Methylomonas, Methylobacter, Alcaligenes, Synechocystis, Anabaena, Thiobacillus, Methanobacterium and Klebsiella.

33 . The non-human organism of claim 31 , wherein said yeast is selected from the group consisting of Aspergillus, Trichoderma, Saccharomyces, Pichia, Candida and Hansenula.

34 . The non-human organism of claim 21 , wherein said organism is an animal.

35 . The non-human organism of claim 34 , wherein said animal is a mammal.

36 . The non-human organism of claim 35 , wherein said mammal is selected from the group consisting of a hamster, a mouse, a rat, a rabbit, a cat, a dog, a bovine, a goat, a pig, a horse and a sheep.

37 . The non-human organism of claim 34 , wherein said animal is an avian.

38 . The non-human organism of claim 37 , wherein said avian is a chicken.

39 . The non-human organism of claim 21 , wherein said organism is a plant.

40 . The non-human organism of claim 39 , wherein said plant is selected from the group consisting of an apple, an Arabidopsis , a bajra, a banana, a barley, a bean, a beet, a blackgram, a chickpea, a chili, a cucumber, an eggplant, a favabean, a maize, a melon, a millet, a mungbean, an oat, an okra, a Panicum , a papaya, a peanut, a pea, a pepper, a pigeonpea, a pineapple, a Phaseolus , a potato, a pumpkin, a rice, a sorghum, a soybean, a squash, a sugarcane, a sugarbeet, a sunflower, a sweet a potato, a tea, a tomato, a tobacco, a watermelon, and a wheat.

41 . The non-human organism of claim 21 , wherein each individually operable heterologous gene modulation system is associated with an expression cassette comprising a response element to which the DNA-binding domain binds, a promoter that is activated by the transactivation domain, and a gene of interest.