Wise/Sost nucleic acid sequences and amino acid sequences

Abstract

The present invention relates to nucleic acid sequences and amino acid sequences which influence bone deposition, the Wnt pathway, ocular development, tooth development, and may bind to LRP. The nucleic acid sequence and polypeptides include Wise and Sost as well as a family of molecules which express a cysteine knot polypeptide. Additionally, the present invention relates to various molecular tools derived from the nucleic acids and polypeptides including vectors, transfected host cells, monochronal antibodies, Fab fragments, and methods for impacting the pathways.

What is claimed is:  
     
         1 . A family of isolated nucleic acid molecules which can influence at least one of the following: tooth development, Wnt pathway activation, bone deposition, or ocular development, wherein the family is selected from the group consisting of: 
 (a) isolated nucleic acid molecule selected from the group consisting of SEQ ID NOs 1-8, 10-28, 96, 97, 108-111, 126, and 127, and complementary sequences thereof;    (b) degenerate variants of the sequences of step a;    (c) an isolated nucleic acid molecule that expresses a cysteine knot protein; and,    (d) oligonucleotide fragments which are 70% homologous to Exon 2 of SEQ. ID. NO. 128.    
     
     
         2 . The isolated nucleic acid molecule of Claim l(c), wherein the nucleic acid molecule is selected from the group consisting of Wise and Sost family member nucleic acid sequence molecules.  
     
     
         3 . The isolated nucleic acid molecule of  claim 1 , wherein nucleic acid molecules are homologous to the sequences of  claim 1 , and are selected from the group consisting of genes, mRNA, cDNA, gDNA, tRNA, RNAi, SiRNA, oligonucleotides, polynucleotides, and nucleic acid sequence fragments.  
     
     
         4 . The isolated nucleic acid molecules of  claim 1  wherein the molecules comprise genes, mRNA, cDNA, gDNA, tRNA, RNAi, oligonucleotides, polynucleotides, and nucleic acid sequence fragments.  
     
     
         5 . Antisense RNAs complementary to at least one of the isolated nucleic acid molecules of  claim 1 .  
     
     
         6 . Mutations of the nucleic acid sequences of  claim 1 , wherein mutants are selected from the group consisting of point, frame shift, deletion, and loss of function mutations.  
     
     
         7 . The mutations of  claim 5  wherein the loss of function mutation comprises a stop codon associated with Exon 1  of the Wise gene.  
     
     
         8 . The mutations of  claim 5 , wherein the loss of function mutation comprises a stop codon associated with the Sost gene.  
     
     
         9 . An antisense oligonucleotide to any mRNA transcribed from at least one nucleic acid molecule of  claim 1 .  
     
     
         10 . An RNAi complementary to at least one of the nucleic acid sequences of  claim 1 .  
     
     
         11 . RNA nucleic acid molecules transcribed from the nucleic acid sequences of  claim 1 .  
     
     
         12 . A probe which hybridizes to at least one of the nucleic acid molecules of  claim 1  selected from the group consisting of cDNA and RNA labeled probes.  
     
     
         13 . A vector comprising a promoter operably linked to a nucleic acid molecule according to  claim 1  or  6 .  
     
     
         14 . The vector of  claim 13 , wherein the vector is selected from the group consisting of expression, cloning, and viral vectors.  
     
     
         15 . The vector of  claim 13 , wherein the vector is selected from the group consisting of expression vectors, fusion vectors, gene therapy vectors, two-hybrid vectors, reverse two-hybrid vectors, sequencing vectors, and cloning vectors.  
     
     
         16 . The vector of  claim 13 , wherein the vector is selected from the group consisting of prokaryotic and eukaryotic vectors.  
     
     
         17 . The prokaryotic vector of  claim 16 , wherein the vector is selected from the group consisting of pET, pET28, pcDNA3.1/V5-His-TOPO, pCS2+, pcDNA II, pSL301, pSE280, pSE380, pSE420, pTrcHis, pRSET, pGEMEX-1, pGEMEX-2, pTrc99A, pKK223-3, pGEX, pEZZ18, pRIT2T, pMC1871, pKK233-2, pKK38801, and pProEx-HT.  
     
     
         18 . The eukaryotic vector of  claim 16 , wherein the vector is selected from the group consisting of pFastBac, pFastBac HT, pFastBac DUAL, pSFV, pTet-Splice, pEUK-C1, pPUR, pMAM, pMAMneo, pBI101, pBI121, pDR2, pCMVEBNA, YACneo, pSVK3, pSVL, pMSG, pCH110, pKK232-8, p3′SS, pBlueBacIII, pCDM8, pcDNA1, pZeoSV, pcDNA3, pREP4, pCEP4, and pEBVHis.  
     
     
         19 . The promoter of  claim 13 , wherein the promotor is selected from the group consisting of a viral promoter and a cellular promoter.  
     
     
         20 . The vector of  claim 13 , wherein the vector comprises a selectable marker selected from the group consisting of an antibiotic resistance gene, a tRNA gene, an auxotrophic gene, a toxic gene, a phenotypic marker, a colorimetric marker, an antisense oligonucleotide, a restriction endonuclease, an enzyme cleavage site, a protein binding site, and an immunoglobulin binding site.  
     
     
         21 . The vector of  claim 20 , wherein the selectable marker is selected from the group consisting of LacZ, neo, Fc, DIG, myc, and FLAG.  
     
     
         22 . The isolated nucleic acid molecule of  claim 1 , wherein nucleic acid molecules homologous to the sequences of  claim 1  are selected from the group consisting of wild type, mutant, antisense, base-substituted, frame shift, deletion, and truncated genes.  
     
     
         23 . The prokaryotic vector of  claim 17 , wherein the vector replicates in a prokaryotic host cell selected from the group consisting of Gram-negative and Gram-positive bacterium.  
     
     
         24 . The prokaryotic host cell of  claim 23 , wherein the host cell is a bacterium selected from the group consisting of Escherichia, Salmonella, Proteus, Clostridium, Klebsiella, Bacillus, Streptomyces, and Pseudomonas.  
     
     
         25 . The Gram-negative bacterium of  claim 23 , wherein the bacterium is  Escherichia coli.    
     
     
         26 . The eukaryotic vector of  claim 16 , wherein the vector replicates in a eukaryotic host cell selected from the group consisting of yeast, plant, fish, mammalian, human, mouse, frog, or insect cells.  
     
     
         27 . The eukaryotic host cell of  claim 26 , wherein the host cell is selected from cells of the group consisting of ES, COS, HEK 293, CHO, SaOS, osteosarcomas, KS483, MG-63, primary osteoblasts, osteoclasts, and human or mammalian bone marrow stroma.  
     
     
         28 . A host cell transfected with a vector according to  claim 13 .  
     
     
         29 . A morpholino antisense oligo molecule derived from any of the nucleic acid sequences of  claim 1 .  
     
     
         30 . The morpholino of  claim 29 , wherein the effective amount of morpholino antisense oligo is within a concentration range between 0.1 nM to 10 mM.  
     
     
         31 . A mutant Wise nucleic acid molecule selected from the group consisting of mutants of the following sequences: 
 (a) isolated nucleic acid molecule selected from the group consisting of SEQ. ID. NOs. 1-5, 96, 97, 109, 126-128, and complementary sequences thereof;    (b) degenerate variants of the sequences of step a;    (c) an isolated nucleic acid molecule that expresses a cysteine knot protein; and,    (d) oligonucleotide fragments which are 70% homologous to Exon 2 of SEQ. ID. NO. 128.    
     
     
         32 . The mutant nucleic acid sequences of  claim 31 , wherein mutants are selected from the group consisting of point, frame shift, deletion, and loss of function mutations.  
     
     
         33 . A recombinant Wise plasmid formed from a mutant of  claim 31 , a promoter, and a selectable marker.  
     
     
         34 . The plasmid of  claim 33  comprising at least one stop codon.  
     
     
         35 . A host cell transfected with the plasmid of  claim 33 , wherein the host cell comprises stem cells.  
     
     
         36 . A host cell transfected with the plasmid of  claim 33 , wherein the host cell comprises embryonic cells.  
     
     
         37 . A chimeric mammal wherein the host cell of  claim 35  or  claim 36  is used to transfect the mammal.  
     
     
         38 . The transfected mammal of  claim 37 , wherein a mouse is selected.  
     
     
         39 . The promoter of  claim 33 , wherein the promotor is selected from the group consisting of a viral promoter and a cellular promoter.  
     
     
         40 . The Wise plasmid of  claim 33 , wherein the selectable marker comprises at least one marker selected from the group consisting of an antibiotic resistance gene, a tRNA gene, an auxotrophic gene, a toxic gene, a phenotypic marker, a colorimetric marker, an antisense oligonucleotide, a restriction endonuclease, an enzyme cleavage site, an enzyme, a protein binding site, and an immunoglobulin binding site.  
     
     
         41 . The Wise plasmid of  claim 33 , wherein the selectable marker is selected from the group consisting of LacZ, neo, Fc, DIG, myc, and FLAG.  
     
     
         42 . The Wise mutant nucleic acid molecule of  claim 31 , wherein the sequences are selected from the group consisting of wild type, mutant, antisense, base-substituted, deletion, frameshift, and truncated genes.  
     
     
         43 . The vector of  claim 33 , wherein the vector is selected from the group consisting of prokaryotic and eukaryotic vectors.  
     
     
         44 . A prokaryotic plasmid of  claim 43 , wherein the plasmid replicates in a prokaryotic host cell selected from the group consisting of Gram-negative and Gram-positive bacterium.  
     
     
         45 . A prokaryotic host cell transfected with the plasmid of  claim 33 , wherein the host cell is selected from the group consisting of Escherichia, Salmonella, Proteus, Clostridium, Klebsiella, Bacillus, Streptomyces, and Pseudomonas.  
     
     
         46 . The Gram-negative bacterium of  claim 43 , wherein the bacterium is  Escherichia coli.    
     
     
         47 . A eukaryotic host cell transfected with the plasmid of  claim 33 , wherein the plasmid replicates in a eukaryotic host cell selected from the group consisting of yeast, plant, fish, mammalian, human, frog, or insect cells.  
     
     
         48 . The eukaryotic host cell of  claim 47 , wherein the host cell is selected from cells of the group consisting of ES, COS, HEK 293, CHO, SaOS, osteosarcomas, KS483, MG-63, primary osteoblasts, osteoclasts, and human or mammalian bone marrow stroma.  
     
     
         49 . A mutant Wise nucleic acid molecule which can influence at least one of the following: tooth development, Wnt pathway, bone deposition, and ocular development selected from the group consisting of: 
 (a) mutants of isolated nucleic acid molecules comprising SEQ. ID. NOs. 1-5, 8, 109, 126-128, and complementary sequences thereof;    (b) degenerate variants of the sequences of step a; and,    (c) Wise nucleic acid molecules having a stop codon which prevents translation to a polypeptide.    
     
     
         50 . A mutant Sost isolated nucleic acid molecule which can influence at least one of the following: tooth development, Wnt pathway, bone deposition, and ocular development selected from the group consisting of mutant variants of: 
 (a) Mutants of isolated nucleic acid molecule comprising SEQ. ID. NOs. 6, 7, 9-14, 110, and 111, and complementary sequences thereof; and,    (b) degenerate variants of the sequences of step a.    
     
     
         51 . The isolated nucleic acid molecule of  claim 50 , wherein nucleic acid molecules are homologous to the sequences of  claim 50 , and are selected from the group consisting of genes, mRNA, cDNA, gDNA, tRNA, RNAi, oligonucleotides, polynucleotides, and nucleic acid sequence fragments.  
     
     
         52 . The mutants of  claim 50 , wherein the mutants comprise antisense RNAs complementary to the non-mutant isolated nucleic acid molecules.  
     
     
         53 . The mutants of  claim 50 , wherein mutants are selected from the group consisting of point, frame shift, deletion, and loss of function mutations.  
     
     
         54 . The mutations of  claim 53 , wherein the loss of function mutation comprises a stop codon at the start of the Sost gene.  
     
     
         55 . An antisense oligonucleotide to any mRNA translated from a nucleic acid molecule of  claim 50 .  
     
     
         56 . An RNAi complementary to the non-mutant nucleic acid sequences homologous to the sequences of  claim 50 .  
     
     
         57 . RNA nucleic acid molecules transcribed from the nucleic acid sequences of  claim 50 .  
     
     
         58 . A probe which hybridizes to at least one of the nucleic acid molecules of  claim 50  selected from the group consisting of cDNA and RNA labeled probes.  
     
     
         59 . A vector comprising a promoter operably linked to a nucleic acid molecule according to  claim 50 .  
     
     
         60 . The vector of  claim 59 , wherein the vector is selected from the group consisting of expression, cloning, and viral vectors.  
     
     
         61 . The vector of  claim 59 , wherein the vector is selected from the group consisting of expression vectors, fusion vectors, gene therapy vectors, two-hybrid vectors, reverse two-hybrid vectors, sequencing vectors, and cloning vectors.  
     
     
         62 . The vector of  claim 59 , wherein the vector is selected from the group consisting of prokaryotic and eukaryotic vectors.  
     
     
         63 . The prokaryotic vector of  claim 62 , wherein the vector is selected from the group consisting of pET, pET28, pcDNA3.1/V5-His-TOPO, pCS2+, pcDNA II, pSL301, pSE280, pSE380, pSE420, pTrcHis, pRSET, pGEMEX-1, pGEMEX-2, pTrc99A, pKK223-3, pGEX, pEZZ18, pRIT2T, pMC1871, pKK233-2, pKK38801, and pProEx-HT.  
     
     
         64 . The eukaryotic vector of  claim 62  wherein the vector is selected from the group consisting of pFastBac, pFastBac HT, pFastBac DUAL, pSFV, pTet-Splice, pEUK-C1, pPUR, pMAM, pMAMneo, pBI101, pBI121, pDR2, pCMVEBNA, YACneo, pSVK3, pSVL, pMSG, pCH110, pKK232-8, p3′SS, pBlueBacIII, pCDM8, pcDNA1, pZeoSV, pcDNA3, pREP4, pCEP4, and pEBVHis.  
     
     
         65 . The promoter of  claim 59 , wherein the promotor is selected from the group consisting of a viral promoter and a cellular promoter.  
     
     
         66 . The vector of  claim 59 , wherein the vector comprises a selectable marker selected from the group consisting of an antibiotic resistance gene, a tRNA gene, an auxotrophic gene, a toxic gene, a phenotypic marker, a calorimetric marker, an antisense oligonucleotide, a restriction endonuclease, an enzyme cleavage site, a protein binding site, and an immunoglobulin binding site.  
     
     
         67 . The vector of  claim 66 , wherein the selectable marker is selected from the group consisting of LacZ, neo, Fc, DIG, myc, and FLAG.  
     
     
         68 . The isolated nucleic acid molecule of  claim 50 , wherein nucleic acid molecules homologous to the sequences of  claim 50  are selected from the group consisting of wild type, mutant, antisense, base-substituted, frame shift, deletion, and truncated genes.  
     
     
         69 . The prokaryotic vector of  claim 63 , wherein the vector replicates in a prokaryotic host cell selected from the group consisting of Gram-negative and Gram-positive bacterium.  
     
     
         70 . The prokaryotic host cell of  claim 69 , wherein the host cell is a bacterium selected from the group consisting of Escherichia, Salmonella, Proteus, Clostridium, Klebsiella, Bacillus, Streptomyces, and Pseudomonas.  
     
     
         71 . The Gram-negative bacterium of  claim 69 , wherein the bacterium is  Escherichia coli.    
     
     
         72 . The eukaryotic vector of  claim 64 , wherein the vector replicates in a eukaryotic host cell selected from the group consisting of yeast, plant, fish, mammalian, human, mouse, frog, or insect cells.  
     
     
         73 . The eukaryotic host cell of  claim 72 , wherein the host cell is selected from cells of the group consisting of ES, COS, HEK 293, CHO, SaOS, osteosarcomas, KS483, MG-63, primary osteoblasts, osteoclasts, and human or mammalian bone marrow stroma.  
     
     
         74 . A host cell transfected with a vector according to  claim 59 .  
     
     
         75 . A morpholino antisense oligo molecule derived from the nucleic acid sequences of  claim 50 .  
     
     
         76 . The morpholino of  claim 75 , wherein the effective amount of morpholino antisense oligo is within a concentration range between 0.1 nM to 10 mM.  
     
     
         77 . A mutant LRP nucleic acid molecule which can influence at least one of the following: tooth development, Wnt pathway activation, bone deposition, or ocular development, selected from the group consisting of: 
 (a) mutants of isolated nucleic acid molecule selected from the group consisting of SEQ ID NOs 29-44, 99, 100, 112, 113, and complementary sequences thereof; and,    (b) degenerate variants of the sequences of step a.    
     
     
         78 . The isolated nucleic acid molecule of  claim 77 , wherein nucleic acid molecules are homologous to the sequences of  claim 77 , and are selected from the group consisting of genes, mRNA, cDNA, gDNA, tRNA, RNAi, oligonucleotides, polynucleotides, and nucleic acid sequence fragments.  
     
     
         79 . The mutants of  claim 77 , wherein the mutants comprise antisense RNAs complementary to the isolated nucleic acid molecules of  claim 77 .  
     
     
         80 . The mutations of  claim 77 , wherein mutants are selected from the group consisting of point, frame shift, deletion, and loss of function mutations.  
     
     
         81 . An antisense oligonucleotide to any mRNA translated from a nucleic acid molecule of  claim 77 .  
     
     
         82 . An RNAi complementary to the nucleic acid sequences homologous to the sequences of  claim 77 .  
     
     
         83 . RNA nucleic acid molecules transcribed from the nucleic acid sequences of  claim 77 .  
     
     
         84 . A probe which hybridizes to at least one of the nucleic acid molecules of  claim 77  selected from the group consisting of cDNA and RNA labeled probes.  
     
     
         85 . A vector comprising a promoter operably linked to a nucleic acid molecule according to  claim 77 .  
     
     
         86 . The vector of  claim 85 , wherein the vector is selected from the group consisting of expression, cloning, and viral vectors.  
     
     
         87 . The vector of  claim 85 , wherein the vector is selected from the group consisting of expression vectors, fusion vectors, gene therapy vectors, two-hybrid vectors, reverse two-hybrid vectors, sequencing vectors, and cloning vectors.  
     
     
         88 . The vector of  claim 85 , wherein the vector is selected from the group consisting of prokaryotic and eukaryotic vectors.  
     
     
         89 . The prokaryotic vector of  claim 88 , wherein the vector is selected from the group consisting of pET, pET28, pcDNA3.1/V5-His-TOPO, pCS2+, pcDNA II, pSL301, pSE280, pSE380, pSE420, pTrcHis, pRSET, pGEMEX-1, pGEMEX-2, pTrc99A, pKK223-3, pGEX, pEZZ18, pRIT2T, pMC1871, pKK233-2, pKK38801, and pProEx-HT.  
     
     
         90 . The eukaryotic vector of  claim 88 , wherein the vector is selected from the group consisting of pFastBac, pFastBac HT, pFastBac DUAL, pSFV, pTet-Splice, pEUK-C1, pPUR, pMAM, pMAMneo, pBI101, pBII21, pDR2, pCMVEBNA, YACneo, pSVK3, pSVL, pMSG, pCH110, pKK232-8, p3′SS, pBlueBacIII, pCDM8, pcDNA1, pZeoSV, pcDNA3, pREP4, pCEP4, and pEBVHis.  
     
     
         91 . The promoter of  claim 85 , wherein the promotor is selected from the group consisting of a viral promoter and a cellular promoter.  
     
     
         92 . The vector of  claim 85 , wherein the vector comprises a selectable marker selected from the group consisting of an antibiotic resistance gene, a tRNA gene, an auxotrophic gene, a toxic gene, a phenotypic marker, a colorimetric marker, an antisense oligonucleotide, a restriction endonuclease, an enzyme cleavage site, a protein binding site, and an immunoglobulin binding site.  
     
     
         93 . The vector of  claim 91 , wherein the selectable marker is selected from the group consisting of LacZ, neo, Fc, DIG, myc, and FLAG.  
     
     
         94 . The isolated nucleic acid molecule of  claim 77 , wherein nucleic acid molecules homologous to the sequences of  claim 77  are selected from the group consisting of wild type, mutant, antisense, base-substituted, frame shift, deletion, and truncated genes.  
     
     
         95 . The prokaryotic vector of  claim 89 , wherein the vector replicates in a prokaryotic host cell selected from the group consisting of Gram-negative and Gram-positive bacterium.  
     
     
         96 . The prokaryotic host cell of  claim 95 , wherein the host cell is a bacterium selected from the group consisting of Escherichia, Salmonella, Proteus, Clostridium, Klebsiella, Bacillus, Streptomyces, and Pseudomonas.  
     
     
         97 . The Gram-negative bacterium of  claim 95 , wherein the bacterium is  Escherichia coli.    
     
     
         98 . The eukaryotic vector of  claim 88 , wherein the vector replicates in a eukaryotic host cell selected from the group consisting of yeast, plant, fish, mammalian, human, mouse, frog, or insect cells.  
     
     
         99 . The eukaryotic host cell of  claim 97 , wherein the host cell is selected from cells of the group consisting of ES, COS, HEK 293, CHO, SaOS, osteosarcomas, KS483, MG-63, primary osteoblasts, osteoclasts, and human or mammalian bone marrow stroma.  
     
     
         100 . A host cell transfected with a vector according to  claim 85 .  
     
     
         101 . A morpholino antisense oligo molecule derived from the nucleic acid sequences of  claim 77 .  
     
     
         102 . The morpholino of  claim 100 , wherein the effective amount of morpholino antisense oligo is within a concentration range between 0.1 nM to 10 mM.  
     
     
         103 . A mutant nucleic acid molecule, wherein the nucleic acid molecule is selected from the group consisting of mutagenized versions of LRP 1, 2, 5, and 6.  
     
     
         104 . A nucleic acid sequence comprising a stop codon and a sequence selected from the group consisting of SEQ. ID. NOs. 1-44, 96-103, 108, 110-113, and 126-128.  
     
     
         105 . A mutant of Wise SEQ. ID. NO. 126.  
     
     
         106 . A mutant of mouse Wise nucleic acid SEQ. ID. NO. 1.  
     
     
         107 . A mutant chick Wise protein SEQ. ID. NO. 4.  
     
     
         108 . A mutant of Wise SEQ. ID. NO. 128.  
     
     
         109 . A mutant of Wise SEQ. ID. NO. 2.  
     
     
         110 . A mutant of Wise SEQ. ID. NO. 96.  
     
     
         111 . A mutant of Wise SEQ. ID. NO. 97.  
     
     
         112 . A mutant of Sost SEQ. ID. NO. 6.  
     
     
         113 . A mutant of Sost SEQ. ID. NO. 8.  
     
     
         114 . A mutant of Sost SEQ. ID. NO. 10.  
     
     
         115 . A mutant of LRP SEQ. ID. NO. 38.  
     
     
         116 . A mutant of LRP SEQ. ID. NO. 39.  
     
     
         117 . A family of amino acid sequences which can influence at least one of the following: tooth development, Wnt pathway activation, bone deposition, or ocular development, selected from the group consisting of: 
 (a) an isolated amino acid sequence comprising SEQ ID NOs 45-48, 50-66, 104-107, 114-125;    (b) Wise amino acid sequences;    (c) Sost amino acid sequences;    (d) LRP amino acid sequences;    (e) an isolated amino acid sequence that is at least 70% homologous, to any of the proteins of (a); and,    (f) an isolated protein that has a cysteine knot formed from eight cysteine residues.    
     
     
         118 . An antibody which binds to at least one of the amino acid sequences of  claim 117 .  
     
     
         119 . The antibodies of  claim 118 , wherein the antibodies are selected from the group consisting of monoclonal antibody, polyclonal antibody, recombinant antibody, and antibody fragment.  
     
     
         120 . A hybridoma cell that expresses at least one the antibodies of  claim 118 .  
     
     
         121 . An antibody that binds to a Wise polypeptide.  
     
     
         122 . An antibody that binds to a Sost polypeptide.  
     
     
         123 . An antibody that selectively binds to an epitope in the receptor-binding domain of the Wise protein.  
     
     
         124 . The antibody of  claim 123 , wherein an epitope on the Wise protein comprises a cysteine knot sequence that binds LRP, wherein the antibody prevents binding of the Wise protein to the LRP.  
     
     
         125 . An Fab fragment derived from an antibody of  claim 118 .  
     
     
         126 . An anti-peptide antibody that prevents binding by Wise amino acid sequences to an LRP polypeptide selected from SEQ. ID. NOs. 67-95.  
     
     
         127 . A Fab fragment that binds to any one of the polypeptides of  claim 117 .  
     
     
         128 . Fab fragments which bind to Exon 2 of Wise.  
     
     
         129 . An anti-peptide antibody that prevents binding by Sost amino acid sequences to an LRP polypeptide selected from SEQ. ID. NOs. 67-95  
     
     
         130 . A family of amino acid sequences selected from the group consisting of: 
 (a) an isolated amino acid sequence comprising SEQ ID NOS 45-53, 104-107, 114-125;    (b) Wise amino acid sequences; and,    (c) SOST amino acid sequences.    
     
     
         131 . An antibody that binds to at least one of the amino acid sequences of  claim 130 .  
     
     
         132 . A Fab fragment from an antibody of  claim 131 .  
     
     
         133 . An isolated amino acid sequence selected from the group consisting of: 
 (a) an isolated amino acid sequence comprising SEQ ID NOS 45, 52, 104, 105, 106, 114-125;    (b) a Wise amino acid sequence encoded by any of the nucleic acid molecules of  claim 1;  and,    (c) an isolated protein that has a cysteine knot formed from eight cysteine residues.    
     
     
         134 . An antibody that binds to at least one of the amino acid sequences of  claim 133 .  
     
     
         135 . A Fab fragment from an antibody of  claim 134 .  
     
     
         136 . A family of amino acid sequences selected from the group consisting of: 
 (a) an isolated amino acid sequence comprising SEQ. ID. NOs. 46-51, 53, 109; and,    (b) a SOST amino acid sequence.    
     
     
         137 . An antibody that binds to at least one of the amino acid sequences of  claim 136 .  
     
     
         138 . A Fab fragment from the antibody of  claim 137 .  
     
     
         139 . An anti-peptide antibody that prevents binding by a Sost amino acid sequences to an LRP polypeptide selected from SEQ. ID. NOs. 67-95.  
     
     
         140 . An isolated amino acid sequence selected from the group consisting of: 
 (a) isolated amino acid sequences comprising SEQ. ID. NOs. 67-95;    (b) LRP polypeptides selected from the group consisting of LRP 1, 2, 5, and 6.    
     
     
         141 . An antibody that binds to at least one of the amino acid sequences of  claim 140 .  
     
     
         142 . A Fab fragment from an antibody of  claim 141 .  
     
     
         143 . An isolated mutant amino acid sequence selected from the group consisting of: 
 (a) isolated mutagenized versions of amino acid sequences selected from SEQ. ID. NOs. 45-48, 50-66, 104-107, 114-125;    (b) mutagenized Wise amino acid sequences; and,    (c) mutagenized Sost amino acid sequences.    
     
     
         144 . An antibody that binds to at least one of the amino acid sequences of  claim 143 .  
     
     
         145 . A Fab fragment from an antibody of  claim 144 .  
     
     
         146 . An isolated antibody derived from the group of polypeptides consisting of: 
 (a) an isolated amino acid sequence comprising SEQ. ID. NOs. 45-95, 104-107, and 114-125;    (b) anti-Wise antibodies;    (c) anti-Sost antibodies; and,    (d) LRP antibodies.    
     
     
         147 . A host cell transfected invitro with an antibody of  claim 146 .  
     
     
         148 . A host cell transfected invivo with an antibody of  claim 146 .  
     
     
         149 . An Fab derived from one of the antibodies of  claim 146 .  
     
     
         150 . An Fab which prevents binding of Sost to LRP where in the Fab is derived from a Sost antibody.  
     
     
         151 . An Fab which prevents binding of Wise to LRP when the Fab is derived from a Wise antibody.  
     
     
         152 . A protein molecule comprising: 
 (a) a Wise polypeptide; and,    (b) an LRP polypeptide.    
     
     
         153 . The protein of  claim 152 , wherein the LRP polypeptide is selected from the group consisting of LRP 1, 2, 5, and 6 polypeptides.  
     
     
         154 . A protein molecule comprising: 
 (a) a Sost polypeptide; and,    (b) an LRP polypeptide.    
     
     
         155 . A method for increasing bone deposition, comprising: 
 (a) isolating a Wise nucleic acid sequence;    (b) attaching a stop codon at the beginning of the Wise nucleic acid sequence to form a Wise cassette;    (c) forming a Wise plasmid by inserting the Wise cassette into the plasmid;    (d) transfecting a host cell with the Wise plasmid, whereby homologous recombination occurs with a wild type Wise gene; and,    (e) activating the stop codon to cause a loss of function mutation.    
     
     
         156 . The method of  claim 155 , wherein the host cell is selected from the group consisting of an insect, an amphibian, and a non-human mammal.  
     
     
         157 . The method of  claim 150 , wherein the host cell is derived from a human.  
     
     
         158 . The method of  claim 155 , wherein expression is controlled by delivery of a Wise nucleic acid molecule into a host cell by a method selected from the group consisting of transfection, microinjection, micro-vessel encapsulation, liposome encapsulation, and electroporation.  
     
     
         159 . The method of  claim 155 , wherein the host cell is selected from the group consisting of osteoblasts and osteoclasts.  
     
     
         160 . The method of  claim 155  comprising transfecting a host organism to form a chimeric host.  
     
     
         161 . The method of  claim 160 , wherein the chimeric host is a mouse.  
     
     
         162 . The method of  claim 160 , wherein the host cells are transfected in vitro.  
     
     
         163 . A method for increasing bone deposition, comprising: 
 (a) isolating a Sost nucleic acid sequence;    (b) attaching a stop code at the beginning of the Sost nucleic acid sequence to form a Sost cassette;    (c) forming a Sost plasmid by inserting the Sost cassette into the plasmid;    (d) transfecting a host cell with the Sost plasmid, whereby homologous recombination occurs with a wild type Sost gene; and,    (e) activating the stop code to cause a loss of function mutation.    
     
     
         164 . The method of  claim 163 , wherein the host cell is selected from the group consisting of an insect, an amphibian, and a non-human mammal.  
     
     
         165 . The method of  claim 163 , wherein the host cell is derived from a human.  
     
     
         166 . The method of  claim 163 , wherein expression is controlled by delivery of a Wise nucleic acid molecule into a host cell by a method selected from the group consisting of microinjection, micro-vessel encapsulation, liposome encapsulation, and electroporation.  
     
     
         167 . The method of  claim 163 , wherein the host cell is selected from the group consisting of osteoblasts and osteoclasts.  
     
     
         168 . The method of  claim 163  comprising transfecting a host organism to form a chimeric host.  
     
     
         169 . The method of  claim 168 , wherein the chimeric host is a mouse.  
     
     
         170 . The method of  claim 163 , wherein the host cells are transfected in vitro.  
     
     
         171 . A method for increasing bone deposition, comprising: 
 (a) isolating a LRP nucleic acid sequence;    (b) attaching a stop code at the beginning of the LRP nucleic acid sequence to form an LRP cassette;    (c) forming an LRP plasmid by inserting the LRP cassette into the plasmid;    (d) transfecting a host cell with the LRP plasmid, whereby homologous recombination occurs with a wild type LRP gene; and,    (e) activating the stop code to cause a loss of function mutation.    
     
     
         172 . The method of  claim 171 , wherein the LRP is selected from the group consisting of LRP 1, 2, 5, and 6.  
     
     
         173 . The method of  claim 171 , wherein expression is controlled by delivery of an LRP nucleic acid molecule into a host cell by a method selected from the group consisting of transfection, microinjection, micro-vessel encapsulation, liposome encapsulation, and electroporation.  
     
     
         174 . The method of  claim 171 , wherein the host cell is selected from the group consisting of osteoblasts and osteoclasts.  
     
     
         175 . The method of  claim 171  comprising transfecting a host organism to form a chimeric host.  
     
     
         176 . The method of  claim 175 , wherein the chimeric host is a mouse.  
     
     
         177 . The method of  claim 171 , wherein the host cells are transfected in vitro.  
     
     
         178 . A method for affecting the Wnt pathway comprising: 
 (a) isolating a Wise nucleic acid sequence;    (b) attaching a stop code at the beginning of the Wise nucleic acid sequence to form a Wise cassette;    (c) forming a Wise plasmid by inserting the Wise cassette into the plasmid;    (d) transfecting a host cell with the Wise plasmid, whereby homologous recombination occurs with a wild type Wise gene; and,    (e) activating the stop code to cause a loss of function mutation.    
     
     
         179 . A method for affecting the Wnt pathway comprising: 
 (a) isolating a Sost nucleic acid sequence;    (b) attaching a stop codon at the beginning of the Sost nucleic acid sequence to form a Sost cassette;    (c) forming a Sost plasmid by inserting the Sost cassette into the plasmid;    (d) transfecting a host cell with the Sost plasmid, whereby homologous recombination occurs with a wild type Sost gene; and,    (e) activating the stop code to cause a loss of function mutation.    
     
     
         180 . A method for affecting the Wnt pathway comprising: 
 (a) isolating an LRP nucleic acid sequence;    (b) attaching a stop codon at the beginning of the LRP nucleic acid sequence to form an LRP cassette;    (c) forming an LRP plasmid by inserting the LRP cassette into the plasmid;    (d) transfecting a host cell with the LRP plasmid, whereby homologous recombination occurs with a wild type LRP gene; and,    (e) activating the stop code to cause a loss of function mutation.    
     
     
         181 . A method for affecting tooth development comprising: 
 (a) isolating a Wise nucleic acid sequence;    (b) attaching a stop code at the beginning of the Wise nucleic acid sequence to form a Wise cassette;    (c) forming a Wise plasmid by inserting the Wise cassette into the plasmid;    (d) transfecting a host cell with the Wise plasmid, whereby homologous recombination occurs with a wild type Wise gene; and,    (e) activating the stop code to cause a loss of function mutation.    
     
     
         182 . A method for affecting tooth development comprising: 
 (a) isolating a Sost nucleic acid sequence;    (b) attaching a stop code at the beginning of the Sost nucleic acid sequence to form a Sost cassette;    (c) forming a Sost plasmid by inserting the Sost cassette into the plasmid;    (d) transfecting a host cell with the Sost plasmid, whereby homologous recombination occurs with a wild type Sost gene; and,    (e) activating the stop code to cause a loss of function mutation.    
     
     
         183 . A method for affecting tooth development comprising: 
 (a) isolating an LRP nucleic acid sequence;    (b) attaching a stop code at the beginning of the LRP nucleic acid sequence to form an LRP cassette;    (c) forming an LRP plasmid by inserting the LRP cassette into the plasmid;    (d) transfecting a host cell with the LRP plasmid, whereby homologous recombination occurs with a wild type LRP gene; and,    (e) activating the stop code to cause a loss of function mutation.    
     
     
         184 . The method of  claim 183 , wherein the plasmid includes a promoter.  
     
     
         185 . The method of  claim 183 , wherein the transfected host cell is delivered to a host organism to form a knockout host.  
     
     
         186 . A method for predicting a defect in bone deposition, comprising: 
 (a) isolating a Wise gene;    (b) forming a labeled Wise gene probe; and,    (c) contacting the labeled gene probe with DNA from a homologue, whereby attachment of the labeled probe indicates a significant probability of normal bone development with normal activation of the Wnt pathway.    
     
     
         187 . A method for causing increased bone deposition comprising: 
 (a) isolating a nucleic acid sequence selected from the group consisting of Wise, Sost, and LRP; and,    (b) forming an antisense RNA from the nucleic acid sequence;    (c) forming an antisense RNA vector; and,    (d) transfecting a host cell with the antisense RNA vector.    
     
     
         188 . The method of  claim 187 , wherein the host cell is selected from the group of animals consisting of insect, amphibian, and non-human mammal.  
     
     
         189 . The method of  claim 187 , wherein the host cell is from  Homo sapiens.    
     
     
         190 . The method of  claim 187 , wherein expression is controlled by injection of an encoding nucleic acid molecule into an embryo.  
     
     
         191 . The method of  claim 187 , wherein the vector is inserted into a prenatal subject.  
     
     
         192 . A method for increasing bone deposition comprising: 
 (a) isolating a Wise nucleic acid sequence;    (b) forming a plasmid vector and transfecting a host cell whereby the host cell expresses the Wise nucleic acid sequence to produce Wise polypeptides;    (c) harvesting the Wise polypeptides;    (d) immunizing a host organism with the Wise polypeptides;    (e) isolating antibodies to the Wise polypeptides from the host;    (f) combining the antibodies with a carrier; and,    (g) transfecting a host cell in vitro.    
     
     
         193 . The method of  claim 192 , wherein a host organism is transfected with the carrier containing the antibodies.  
     
     
         194 . The method of  claim 192 , wherein the host cell is transfected in vivo.  
     
     
         195 . The method of  claim 192 , wherein expression is controlled by delivery of a Wise nucleic acid molecule into a host cell by a method selected from the group consisting of transfection, microinjection, micro-vessel encapsulation, liposome encapsulation, and electroporation.  
     
     
         196 . The method of  claim 192 , wherein the host cell is selected from the group consisting of osteoblasts and osteoclasts.  
     
     
         197 . The method of  claim 192 , wherein the antibodies are Fab fragments.  
     
     
         198 . A method for increasing bone deposition comprising: 
 (a) isolating a Sost nucleic acid sequence;    (b) forming a plasmid vector and transfecting a host cell whereby the host cell expresses the Sost nucleic acid sequence to produce Sost polypeptides;    (c) immunizing a host organism with the Sost polypeptides;    (d) isolating antibodies to the Sost polypeptides from the host;    (e) combining the antibodies with a carrier; and,    (f) transfecting a host cell in vitro.    
     
     
         199 . A method for increasing bone deposition comprising: 
 (a) isolating an LRP nucleic acid sequence;    (b) forming a plasmid vector and transfecting a host cell whereby the host cell expresses the LRP nucleic acid sequence to produce LRP polypeptides;    (c) immunizing a host organism with the LRP polypeptides;    (d) isolating antibodies to the LRP polypeptides from the host;    (e) combining the antibodies with a carrier; and,    (f) transfecting a host cell in vitro.    
     
     
         200 . A method for affecting the Wnt pathway comprising: 
 (a) isolating a Wise nucleic acid sequence;    (b) forming a plasmid vector and transfecting a host cell whereby the host cell expresses the Wise nucleic acid sequence to produce Wise polypeptides;    (c) immunizing a host organism with the Wise polypeptides;    (d) isolating antibodies to the Wise polypeptides from the host;    (e) combining the antibodies with a carrier; and,    (f) transfecting a host cell in vitro.    
     
     
         201 . A method for affecting the Wnt pathway comprising: 
 (a) isolating a Sost nucleic acid sequence;    (b) forming a plasmid vector and transfecting a host cell whereby the host cell expresses the Sost nucleic acid sequence to produce Sost polypeptides;    (c) immunizing a host organism with the Sost polypeptides;    (d) isolating antibodies to the Sost polypeptides from the host;    (e) combining the antibodies with a carrier; and,    (f) transfecting a host cell in vitro.    
     
     
         202 . A method for affecting the Wnt pathway comprising: 
 (a) isolating an LRP nucleic acid sequence;    (b) forming a plasmid vector and transfecting a host cell whereby the host cell expresses the LRP nucleic acid sequence to produce LRP polypeptides;    (c) immunizing a host organism with the LRP polypeptides;    (d) isolating Fab antibodies to the LRP polypeptides from the host;    (e) combining the antibodies with a carrier; and,    (f) transfecting a host cell in vitro.    
     
     
         203 . A method for affecting tooth development comprising: 
 (a) isolating a nucleic acid sequence selected from the group consisting of Wise, Sost, and LRP;    (b) forming a plasmid vector and transfecting a host cell whereby the host cell expresses the nucleic acid sequence to produce polypeptides;    (c) immunizing a host organism with the polypeptides;    (d) isolating antibodies to the polypeptides from the host;    (e) combining the antibodies with a carrier; and,    (f) transfecting a host cell in vivo.    
     
     
         204 . A method for affecting ocular development comprising: 
 (a) isolating a nucleic acid sequence selected from the group consisting of Wise, Sost, and LRP;    (b) forming a plasmid vector and transfecting a host cell whereby the host cell expresses the nucleic acid sequence to produce polypeptides;    (c) immunizing a host organism with the polypeptides;    (d) isolating antibodies to the polypeptides from the host;    (e) combining the antibodies with a carrier; and,    (f) transfecting a host cell in vivo.    
     
     
         205 . A method for preventing Sost from binding to an LRP selected from the group consisting of LRP5 and LRP6 comprising: 
 (a) isolating a Sost nucleic acid sequence;    (b) forming a plasmid vector and transfecting a host cell whereby the host cell expresses the Sost nucleic acid sequence to produce Sost polypeptides;    (c) immunizing a host organism with the Sost polypeptides;    (d) isolating antibodies to the LRP polypeptides from the host;    (e) combining the antibodies with a carrier; and,    (f) transfecting a host cell in vivo.    
     
     
         206 . A method for preventing Wise from binding to an LRP selected from the group consisting of LRP5 and LRP6 comprising: 
 (a) isolating a Wise nucleic acid sequence;    (b) forming a plasmid vector and transfecting a host cell whereby the host cell expresses the Wise nucleic acid sequence to produce Wise polypeptides;    (c) immunizing a host organism with the Wise polypeptides;    (d) isolating antibodies to the Wise polypeptides from the host;    (e) combining the antibodies with a carrier; and,    (f) transfecting a host cell in vitro.    
     
     
         207 . A method for affecting either bone deposition, ocular development, Wnt pathway, or tooth development, comprising transfecting a host cell with an antibody derived from the group consisting of antibodies to LRP, Wise, and Sost, wherein the antibody prevents wild-type polypeptides from binding with their targets.  
     
     
         208 . A kit for detecting a Wise polypeptide, wherein the kit comprises: 
 (a) a container; and,    (b) a Wise antibody with a marker.    
     
     
         209 . A kit for detecting a Wise nucleic acid molecule, wherein the kit comprises: 
 (a) a container; and,    (b) a Wise probe.    
     
     
         210 . The method of  claim 207 , wherein the protein molecule is isolated from a host organism in the group selected from Humans, xenopus, frogs, and Drosophila.  
     
     
         211 . A method for blocking Wise/SOST expression using a morpholino.  
     
     
         212 . The kit of  claim 208 , wherein the markers comprise en2, Krox20, Hoxb9, myosin, RT-PCR, Ef1-Δ, NCAM, otx2, myosin light chain, and muscle actin.  
     
     
         213 . The method of  claim 208 , wherein the markers are selected from the group consisting of posterior, midbrain, hindbrain, spinal cord, mesoderm, muscle, and neural markers.  
     
     
         214 . A Wise nucleic acid sequence comprising Wise, hemaglutinin, myc, stop codon, and FLAG sequences.  
     
     
         215 . A method for activating canonical Wnt signaling comprising: 
 (a) injecting a Wise protein into an embryo, wherein the Wise protein binds to a Frizzled receptor thereby inhibiting the binding of the Wnt protein to the Frizzled receptor; and,    (b) activating canonical Wnt signaling.    
     
     
         216 . A family of nucleic acid sequences selected from the group consisting of Caronte, Wise, Sost Dan, Cereberus, Gremlin, CTGF, Soggy, DKK1, DKK2, DKK3, DKK4, NOV, mucin, slit, OH, WISP, and CCN.  
     
     
         217 . A method for producing a Wise mutant mouse comprising: 
 (a) introducing a mutant Wise gene into a mouse embryonic stem cell;    (b) introducing the embryonic stem cell into a mouse blastocyst to create a transgenic embryo; and,    (c) allowing the embryo to develop into the Wise mouse.    
     
     
         218 . The method of  claim 217 , wherein the introduction of a gene into the stem cell is selected from the group of methods consisting of transfection, micro-injection, biolistic particle delivery, lipofection, and electroporation.  
     
     
         219 . The method of  claim 217 , wherein the Wise mouse exhibits developmental abnormalities selected from the group consisting of bone deposition, dental, neurological, and ocular abnormalities.  
     
     
         220 . The mutated Wise gene of  claim 217 , wherein the gene is selected from the group consisting of antisense, base-substituted, and truncated gene sequences.  
     
     
         221 . An isolated mouse cell comprising a mutated Wise gene, wherein the endogenous wild type Wise gene has been replaced with the mutated Wise gene.  
     
     
         222 . The mutated Wise gene of  claim 221 , wherein the gene is selected from the group consisting of antisense, base-substituted, and truncated genes.  
     
     
         223 . A Wise pET vector comprising a mutated Wise gene sequence, neo, and LacZ.  
     
     
         224 . The mutated Wise sequence of  claim 223 , wherein the gene sequence is selected from the group consisting of antisense, base-substituted, and truncated sequences.  
     
     
         225 . A Sost pET vector comprising a mutated Sost sequence, neo, and LacZ.  
     
     
         226 . The mutated Sost sequence of  claim 225 , wherein the gene sequence is selected from the group consisting of antisense, base-substituted, and truncated sequences.  
     
     
         227 . A mutant Wise mouse comprising the mutant Wise nucleic acid sequence of  claim 31 .  
     
     
         228 . The Wise mouse of  claim 227 , wherein the mutant Wise gene is selected from the group consisting of homozygous and heterozygous genes.  
     
     
         229 . A Sost mouse comprising the mutant Sost gene sequence of  claim 49 .  
     
     
         230 . The Sost mouse of  claim 229 , wherein the mutant Sost gene is selected from the group consisting of homozygous and heterozygous genes.  
     
     
         231 . A mutant Wise mouse made by the steps comprising: 
 (a) introducing a mutant Wise gene into a mouse embryonic stem cell;    (b) introducing the embryonic stem cell into a mouse blastocyst to create a transgenic embryo; and,    (c) allowing the embryo to develop into the mutant Wise mouse.    
     
     
         232 . A family of nucleic acid sequences which can influence at least one of the following: bone deposition, Wnt pathway, tooth development, and ocular development, selected from the group consisting of SEQ. ID. NOs. 1-44.  
     
     
         233 . An isolated nucleic acid sequence which regulates bone deposition, ocular development, Wnt pathway, and tooth development, and binds to LRP.  
     
     
         234 . The isolated nucleic acid sequence of  claim 233 , wherein the sequence is selected from the group consisting of Wise and Sost nucleic acid sequences.  
     
     
         235 . A method for producing a transgenic mutant Wise mouse, comprising: 
 (a) microinjecting a Wise cassette containing a mutant Wise nucleic acid sequence into a blastomere;    (b) injecting the blastomere into a host mouse embryo; and,    (c) growing the embryo to maturation.