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1.发明公开
公开(公告)号：EP1507865A2
公开(公告)日：2005-02-23
申请号：EP03732512.3
申请日：2003-05-30
CPC分类号： C12N15/90 , C12N15/85 , C12N2800/60 , C12N2800/90 , C12N2840/44
摘要： The present invention relates to a transposon-based DNA integration system comprising (a) a transposon which is devoid of a polynucleotide encoding a functional transposase and which comprises a polynucleotide of interest, wherein the transposon comprises inverted repeats having a degree of identity with the repeats within SEQ ID NO: 2 and its inverted repeat, respectively, of at least 90%; and (b) a transposase having at its N-terminus a DNA binding domain comprising the sequences of SEQ ID NO: 3 and 4; or (c) a polynucleotide encoding the transposase of (b). The present invention further relates to a method of transferring a polynucleotide of interest into cells of a vertebrate comprising the step of introducing the transposon-based DNA integration system of the invention into said cells. In addition, the invention relates to a method of effecting RNAi comprising (a) stably introducing a transposon comprising an expression cassette expressing a short interfering RNA and a selectable marker gene as part of the trans poson-based DNA integration system of the invention into a cell; (b) selecting for cells expressing the selectable marker; and (c) assessing whether the transcription/translation of the desired gene is effected by RNAi. A further embodiment of the invention is a method of gene trapping genes comprising the steps of (a) introducing the transposon-based DNA integration system of the invention into a cell; and (b) assessing for the expression of a selectable marker wherein expression of a selectable marker is indicative of integration of the transposon into a transcibed gene of the cell.
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2.发明授权
公开(公告)号：EP1507865B1
公开(公告)日：2008-07-16
申请号：EP03732512.3
申请日：2003-05-30
CPC分类号： C12N15/90 , C12N15/85 , C12N2800/60 , C12N2800/90 , C12N2840/44
摘要： The present invention relates to a transposon-based DNA integration system comprising (a) a transposon which is devoid of a polynucleotide encoding a functional transposase and which comprises a polynucleotide of interest, wherein the transposon comprises inverted repeats having a degree of identity with the repeats within SEQ ID NO: 2 and its inverted repeat, respectively, of at least 90%; and (b) a transposase having at its N-terminus a DNA binding domain comprising the sequences of SEQ ID NO: 3 and 4; or (c) a polynucleotide encoding the transposase of (b). The present invention further relates to a method of transferring a polynucleotide of interest into cells of a vertebrate comprising the step of introducing the transposon-based DNA integration system of the invention into said cells. In addition, the invention relates to a method of effecting RNAi comprising (a) stably introducing a transposon comprising an expression cassette expressing a short interfering RNA and a selectable marker gene as part of the trans poson-based DNA integration system of the invention into a cell; (b) selecting for cells expressing the selectable marker; and (c) assessing whether the transcription/translation of the desired gene is effected by RNAi. A further embodiment of the invention is a method of gene trapping genes comprising the steps of (a) introducing the transposon-based DNA integration system of the invention into a cell; and (b) assessing for the expression of a selectable marker wherein expression of a selectable marker is indicative of integration of the transposon into a transcibed gene of the cell.
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3.发明公开HYPERACTIVE VARIANTS OF THE TRANSPOSASE PROTEIN OF THE TRANSPOSON SYSTEM SLEEPING BEAUTY 有权多动版DES TRANSPOSASEPROTEINS DES睡袋美容TRANSPOSONSYSTEMS
公开(公告)号：EP2160461A2
公开(公告)日：2010-03-10
申请号：EP08773780.5
申请日：2008-06-30
发明人： IZSVAK, Zsuzsanna , IVICS, Zoltan , MATES, Lajos , MANOJ, Namitha , JUDIS, Carmen-Anisia , KATZER, Andrea
CPC分类号： C12N9/1241 , A61K38/45 , A61K48/00 , A61K48/0066 , C12N9/22 , C12N15/90 , C12N2800/90 , C12Y207/07
摘要： The present invention refers to hyperactive variants of a transposase of the transposon system Sleeping Beauty (SB). The invention further refers to corresponding nucleic acids producing these variants, to a gene transfer system for stably introducing nucleic acid(s) into the DNA of a cell by using these hyperactive variants of a transposase of the transposon system Sleeping Beauty (SB) and to transposons used in the inventive gene transfer system, comprising a nucleic acid sequence with flanking repeats (IRs and/or RSDs). Furthermore, applications of these transposase variants, the transpsoson, or the gene transfer system are also disclosed such as gene therapy, insertional mutagenesis, gene discovery (including genome mapping), mobilization of genes, library screening, or functional analysis of genomes in vivo and in vitro. Finally, pharmaceutical compositions and kits are also encompassed.
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公开(公告)号：EP1594972A2
公开(公告)日：2005-11-16
申请号：EP04709603.7
申请日：2004-02-10
发明人： IVICS, Zoltán , IZSVAK, Zsuzsanna
CPC分类号： C12N15/90
摘要： The present invention relates to a targeting system comprising, preferably as distinct components, (a) a transposon which is devoid of a polynucleotide encoding a functional transposase comprising a polynucleotide of interest; and (b) a fusion protein comprising (ba) a transposase or a fragment or derivative thereof having transposase function; and (bb) a DNA targeting domain; or (bc) a (poly)peptide binding domain that binds to a cellular or engineered (poly)peptide comprising a DNA targeting domain; or (bd) a (poly)peptide comprising the DNA targeting domain of (bb) or the (poly)peptide binding domain of (bc), wherein the transposase or a fragment or derivative thereof having transposase function of (a) is joined by a linker to the domain of (bb) or to the domain of (bc) or to the (poly)peptide of (bd); or (c) a polynucleotide encoding the fusion protein of (b).
摘要翻译： 本发明涉及一种靶向系统，其优选包含（a）不含编码包含目的多核苷酸的功能性转座酶的多核苷酸的转座子; 和（b）融合蛋白，其包含（ba）具有转座酶功能的转座酶或其片段或衍生物; 和（bb）DNA靶向结构域; 或（bc）结合包含DNA靶向性结构域的细胞或工程化（多）肽的（多）肽结合结构域; 或（bd）包含（bb）的DNA靶向性结构域或（bc）的（多）肽结合性结构域的（多）肽，其中具有（a）的转座酶功能的转座酶或其片段或衍生物通过 （bb）的结构域或（bc）的结构域或（bd）的（多）肽的结构域的接头; 或（c）编码（b）的融合蛋白的多核苷酸。
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公开(公告)号：EP1594972B1
公开(公告)日：2011-02-09
申请号：EP04709603.7
申请日：2004-02-10
发明人： IVICS, Zoltán , IZSVAK, Zsuzsanna
CPC分类号： C12N15/90
摘要： The present invention relates to a targeting system comprising, preferably as distinct components, (a) a transposon which is devoid of a polynucleotide encoding a functional transposase comprising a polynucleotide of interest; and (b) a fusion protein comprising (ba) a transposase or a fragment or derivative thereof having transposase function; and (bb) a DNA targeting domain; or (bc) a (poly)peptide binding domain that binds to a cellular or engineered (poly)peptide comprising a DNA targeting domain; or (bd) a (poly)peptide comprising the DNA targeting domain of (bb) or the (poly)peptide binding domain of (bc), wherein the transposase or a fragment or derivative thereof having transposase function of (a) is joined by a linker to the domain of (bb) or to the domain of (bc) or to the (poly)peptide of (bd); or (c) a polynucleotide encoding the fusion protein of (b).
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公开(公告)号：EP1594973A1
公开(公告)日：2005-11-16
申请号：EP04709610.2
申请日：2004-02-10
发明人： IVICS, Zoltán , IZSVAK, Zsuzsanna
CPC分类号： C12N15/90
摘要： The present invention relates to a targeting system comprising, preferably as distinct components (a) a transposon which is devoid of polynucleotide encoding a functional transposase comprising (aa) a polynucleotide of interest and; (ab) a DNA sequence specifically recognized by a DNA binding domain; and (ba) a fusion protein comprising (i) said DNA binding domain; or (ii) a (poly)peptide binding domain binding to a (poly)peptide comprising said DNA binding domain; and (iii) a DNA targeting domain; or (iv) a (poly)peptide binding domain that binds to a cellular or engineered (poly)peptide that comprises a DNA targeting domain; or (bb) a polynucleotide encoding the fusion protein of (ba); and (ca) a transposase or a fragment or derivative thereof having transposase function; or (cb) a polynucleotide encoding the transposase or fragment or derivative thereof having transposase function of (ca).
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7.发明授权
公开(公告)号：EP1276889B1
公开(公告)日：2005-10-19
申请号：EP01942975.2
申请日：2001-04-27
发明人： IVICS, Zoltan , IZSVAK, Zsuzsanna , Zayed, Hatem
IPC分类号： C12N15/90 , C12N15/67 , C12N5/16 , A01K67/027
CPC分类号： C12N15/90 , A01K2217/05
摘要： The invention relates to the use of the gene transfer system Sleeping Beauty for the somatic gene transfer for the purpose of stably inserting DNA in the chromosomes of living vertebrates, comprising the two components of the transfer system Sleeping Beauty that are injected into the somatic cells of an animal for the purpose of gene therapy.
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8.发明公开
公开(公告)号：EP1276889A2
公开(公告)日：2003-01-22
申请号：EP01942975.2
申请日：2001-04-27
发明人： IVICS, Zoltan , IZSVAK, Zsuzsanna
IPC分类号： C12N15/90 , C12N15/67 , C12N5/16 , A01K67/027
CPC分类号： C12N15/90 , A01K2217/05
摘要： The invention relates to the use of the gene transfer system Sleeping Beauty for the somatic gene transfer for the purpose of stably inserting DNA in the chromosomes of living vertebrates, comprising the two components of the transfer system Sleeping Beauty that are injected into the somatic cells of an animal for the purpose of gene therapy.
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9.发明授权HYPERACTIVE VARIANTS OF THE TRANSPOSASE PROTEIN OF THE TRANSPOSON SYSTEM SLEEPING BEAUTY 有权TRANSPOSASEPROTEINS的的过度活动类型睡美人，转座子
公开(公告)号：EP2160461B1
公开(公告)日：2012-08-08
申请号：EP08773780.5
申请日：2008-06-30
发明人： IZSVAK, Zsuzsanna , IVICS, Zoltan , MATES, Lajos , MANOJ, Namitha , JUDIS, Carmen-Anisia , KATZER, Andrea
CPC分类号： C12N9/1241 , A61K38/45 , A61K48/00 , A61K48/0066 , C12N9/22 , C12N15/90 , C12N2800/90 , C12Y207/07
摘要： The present invention refers to hyperactive variants of a transposase of the transposon system Sleeping Beauty (SB). The invention further refers to corresponding nucleic acids producing these variants, to a gene transfer system for stably introducing nucleic acid(s) into the DNA of a cell by using these hyperactive variants of a transposase of the transposon system Sleeping Beauty (SB) and to transposons used in the inventive gene transfer system, comprising a nucleic acid sequence with flanking repeats (IRs and/or RSDs). Furthermore, applications of these transposase variants, the transpsoson, or the gene transfer system are also disclosed such as gene therapy, insertional mutagenesis, gene discovery (including genome mapping), mobilization of genes, library screening, or functional analysis of genomes in vivo and in vitro. Finally, pharmaceutical compositions and kits are also encompassed.
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公开(公告)号：EP1594973B1
公开(公告)日：2011-12-07
申请号：EP04709610.2
申请日：2004-02-10
发明人： IVICS, Zoltán , IZSVAK, Zsuzsanna
CPC分类号： C12N15/90
摘要： The present invention relates to a targeting system comprising, preferably as distinct components (a) a transposon which is devoid of polynucleotide encoding a functional transposase comprising (aa) a polynucleotide of interest and; (ab) a DNA sequence specifically recognized by a DNA binding domain; and (ba) a fusion protein comprising (i) said DNA binding domain; or (ii) a (poly)peptide binding domain binding to a (poly)peptide comprising said DNA binding domain; and (iii) a DNA targeting domain; or (iv) a (poly)peptide binding domain that binds to a cellular or engineered (poly)peptide that comprises a DNA targeting domain; or (bb) a polynucleotide encoding the fusion protein of (ba); and (ca) a transposase or a fragment or derivative thereof having transposase function; or (cb) a polynucleotide encoding the transposase or fragment or derivative thereof having transposase function of (ca).
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