Transgenic animals with customizable traits

Abstract

Disclosed are materials and methods for creating customizable traits in animals. In the demonstration of the principle of the subject invention, a keratin-14 specific promoter is used with, red fluorescent protein in the loxp cassette, dominant black (ΔG23) beta defensin 103 in the pigment cassette, and an SV40 (with intron) polyadenylation sequence. When Cre recombinase (or HTNCre) is applied to the animal's skin in a carrier base (e.g., lipid bilayers), fur is permanently genetically modified to turn black in the shape in which it was applied.

I claim: 
     
         1 . A cell of a transgenic non-human animal, wherein the cell comprises:
 an exogenous nucleic acid molecule operably linked to a promoter, wherein the expression of the exogenous nucleic acid molecule is under the control of an inducible expression system that requires the presence of an inducing agent to activate expression; and   wherein the expression of the exogenous nucleic acid molecule is inhibited in the absence of the inducing agent;   wherein the exogenous nucleic acid molecule is selected from:   a nucleic acid molecule encoding a protein selected from pigment proteins; proteins involved in the synthesis and/or transport of biological pigments; proteins involved in skin texture; proteins involved in the fur texture and/or length; and proteins involved in the texture, structural strength, and/or length of nail, claw, and/or horn; and   an inhibitory RNA coding sequence that interferes with the expression of a nucleic acid molecule encoding a protein selected from pigment proteins; proteins involved in the synthesis and/or transport of biological pigments; proteins involved in skin texture; proteins involved in the fur texture and/or length; and proteins involved in the texture, structural strength, and/or length of nail, claw, and/or horn.   
     
     
         2 . A non-human transgenic animal comprising a cell according to  claim 1 . 
     
     
         3 . The non-human transgenic animal according to  claim 2 , wherein the inducible gene expression system is selected from a Cre-LoxP recombination system, a FLP-FRT recombination system, a tetracycline (Tet)-controlled transcription activation system, an ecdysone inducible system, a heat shock on/off system, a lacO/IPTG system, a cumate repressor protein CymR system, a nitroreductase system, coumermycin/novobiocin-regulated system, a RheoSwitch Ligand RSL/system, a chimeric bipartite nuclear receptor expression system, a GAL4 system, sterol or steroid or synthetic steroid inducing/repressing system, and any combination thereof. 
     
     
         4 . The non-human transgenic animal according to  claim 3 , wherein the inducible gene expression system comprises a Cre-LoxP recombination system. 
     
     
         5 . The non-human transgenic animal according to  claim 4 , wherein the Cre-LoxP recombination system comprises a lox-stop-lox (LSL) sequence. 
     
     
         6 . The non-human transgenic animal according to  claim 4 , wherein the inducing agent is a Cre recombinase protein. 
     
     
         7 . The non-human transgenic animal according to  claim 3 , wherein the inducible gene expression system comprises a tetracycline (Tet)-controlled transcription activation system. 
     
     
         8 . The non-human transgenic animal according to  claim 7 , wherein the inducing agent is doxycycline. 
     
     
         9 . The non-human transgenic animal according to  claim 2 , wherein the promoter is selected from universal promoters, constitutive promoters, tissue-specific promoters, and inducible promoters. 
     
     
         10 . The non-human transgenic animal according to  claim 2 , wherein the promoter is selected from cytomegalovirus (CMV) promoter, CMV-chicken beta actin promoter, ubiquitin promoter, JeT promoter, SV40 promoter, beta globin promoter, elongation Factor 1 alpha (EF1-alpha) promoter, RSV promoter, Ripply2 promoter, Ticked promoter, Tabby promoter, Mo-MLV-LTR promoter,  Rosa 26 promoter, keratinocyte specific promoters, melanocyte specific promoters, matrix-cell specific promoters, and dermal papilla-specific promoters. 
     
     
         11 . The non-human transgenic animal according to  claim 2 , comprising a cell that comprises:
 a first exogenous nucleic acid molecule operably linked to a first promoter and under the control of a loxP site, wherein the loxP site prevents the expression of the first nucleic acid molecule in the absence of Cre recombinase protein;   a second exogenous nucleic acid molecule encoding a reverse tTA (rtTA), operably linked to a second promoter; and   a third exogenous nucleic acid molecule encoding a Cre recombinase protein, operably linked to a third promoter under the control of a TetO operator;   wherein the first exogenous nucleic acid molecule is selected from:   a nucleic acid molecule encoding a protein selected from pigment proteins; proteins involved in the synthesis and/or transport of biological pigments; proteins involved in skin texture; proteins involved in the fur texture and/or length; and proteins involved in the texture, structural strength, and/or length of nail, claw, and/or horn; and   an inhibitory RNA coding sequence that interferes with the expression of a nucleic acid molecule encoding a protein selected from pigment proteins; proteins involved in the synthesis and/or transport of biological pigments; proteins involved in skin texture; proteins involved in the fur texture and/or length; and proteins involved in the texture, structural strength, and/or length of nail, claw, and/or horn.   
     
     
         12 . The non-human transgenic animal according to  claim 2 , wherein the exogenous nucleic acid molecule encodes a protein selected from pigment proteins; proteins involved in the synthesis and/or transport of biological pigments; proteins involved in skin texture; proteins involved in the fur texture and/or length; and proteins involved in the texture, structural strength, and/or length of nail, claw, and/or horn. 
     
     
         13 . The non-human transgenic animal according to  claim 12 , wherein the exogenous nucleic acid molecule encodes a protein selected from melanocortin receptor (MC1R), melanocyte stimulating hormones (MSH), β-defensin 103, agouti signaling protein (ASP), tyrosinase (TYR), melanocyte-specific transporter protein, Ras-related protein Rab-7, rab protein geranylgeranyltransferase component A2, probable E3 ubiquitin-protein ligase (HERC2), bifunctional enzyme CarRP-like, lycopene cyclase/phytoene synthase-like, and phytoene dehydrogenase-like. 
     
     
         14 . The non-human transgenic animal according to  claim 2 , wherein the exogenous nucleic acid molecule comprises an inhibitory RNA coding sequence that interferes with the expression of a nucleic acid molecule encoding a protein selected from pigment proteins; proteins involved in the synthesis and/or transport of biological pigments; proteins involved in skin texture; proteins involved in the fur texture and/or length; and proteins involved in the texture, structural strength, and/or length of nail, claw, and/or horn. 
     
     
         15 . The non-human transgenic animal according to  claim 14 , wherein the inhibitory RNA coding sequence interferes with the expression of a nucleic acid molecule encoding a keratin protein. 
     
     
         16 . The non-human transgenic animal according to  claim 2 , which is a dog, cat, mouse, rat, guinea pig, hamster, horse, cow, pig, sheep, goat, duck, goose, chicken, primate, fish, frog, salamander, snake, lizard, fox, weasel, rabbit, mink, beaver, ermine, otter, sable, seal, coyote, chinchilla, deer, muskrat, or possum. 
     
     
         17 . A method of customizing non-human animal traits comprising:
 providing a non-human transgenic animal according to  claim 2 ;   administering the inducing agent to the transgenic animal thereby inducing the expression of the exogenous nucleic acid molecule.   
     
     
         18 . The method according to  claim 17 , wherein the inducing agent is formulated with a carrier for administration to the transgenic animal. 
     
     
         19 . The method according to  claim 17 , wherein the inducible gene expression system comprises a Cre-LoxP recombination system, the inducing agent is a Cre recombinase protein, and the Cre recombinase is topically administered to the transgenic animal. 
     
     
         20 . The method according to  claim 17 , wherein the exogenous nucleic acid molecule is transferred into the animal via the use of spermatogonial stem cells (SSCs), piggyBac™ mobile DNA technology using transposable elements,  Xanthamonas  transcription activator-like (TAL) Nucleases (XTNs), or a combination thereof.