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1. 发明申请

WO1998011214A1 MAMMALIAN AND HUMAN REC2 审中-公开
标题翻译：妈妈和人类REC2
公开(公告)号：WO1998011214A1
公开(公告)日：1998-03-19
申请号：PCT/IB1997001217
申请日：1997-09-11
申请人： THOMAS JEFFERSON UNIVERSITY , CORNELL RESEARCH FOUNDATION, INC.
发明人： THOMAS JEFFERSON UNIVERSITY , CORNELL RESEARCH FOUNDATION, INC. , HOLLOMAN, William, K. , RICE, Michael, C. , SMITH, Sheryl, T. , SHU, Zhigang , KMIEC, Eric, B.
IPC分类号： C12N15/12
CPC分类号： C12N9/00 , A01K2217/05
摘要： The invention concerns mammalian recombinase genes (REC2) and their promoters. Over expression of REC2 in a cell is found to facilitate homologous recombination, particularly homologous recombination using a DNA/RNA chimeric oligonucleotide and to sensitize a cell to the apoptotic effects of irradiation. The REC2 promoter, in combination with a strong enhancer, e.g., a SV40 enhancer, was found to be a strong promoter following irridiation of the cells. A radiation induceable promoter can be used to sensitize a cell to radiation treatment by operably linking the radiation induceable promoter to a gene whose expression converts a prodrug to a drug such as a herpes thymidine kinase gene.
摘要翻译：本发明涉及哺乳动物重组酶基因（REC2）及其启动子。发现细胞中REC2的过表达促进同源重组，特别是使用DNA / RNA嵌合寡核苷酸的同源重组，并使细胞对辐照的凋亡作用敏感。发现REC2启动子与强增强子（例如SV40增强子）组合，在细胞灌注后是强启动子。辐射诱导启动子可用于通过将辐射诱导启动子可操作地连接到表达将前药转化为药物如疱疹胸苷激酶基因的基因来使细胞对辐射治疗敏感。

2. 发明申请

WO1996022364A1 COMPOSITIONS AND METHODS TO PROMOTE HOMOLOGOUS RECOMBINATION IN EUKARYOTIC CELLS AND ORGANISMS 审中-公开
标题翻译：促进真核细胞和有机体中的同源重组的组合物和方法
公开(公告)号：WO1996022364A1
公开(公告)日：1996-07-25
申请号：PCT/US1996000265
申请日：1996-01-16
申请人： CORNELL RESEARCH FOUNDATION, INC. , THOMAS JEFFERSON UNIVERSITY
发明人： CORNELL RESEARCH FOUNDATION, INC. , THOMAS JEFFERSON UNIVERSITY , HOLLOMAN, William, K. , KMIEC, Eric, B.
IPC分类号： C12N09/00
CPC分类号： C12N9/00
摘要： The invention concerns genes encoding recombinases that can be used to promote homologous recombination in eukaryotic cells. The application teaches methods by which a recombinase of one species can be used to isolate a homologous recombinase of a different species and methods to identify the isolated homologs. Recombinases from Ustilago maydis and Saccharomyces cerevisiae are specifically included in the invention. The invention encompasses the method of producing an isolated recombinase protein in a prokaryotic cell and recovering the product in an active form. The invention also encompasses a genetically engineered gene which encodes a non-naturally occurring recombinase that causes a greater rate of recombination than does the naturally occurring recombinase. The invention further encompasses the use of recombinase proteins and of recombinase genes to promote homologous recombination, including recombination between a host cell genome and a chimeric oligonucleotide, i.e., an oligonucleotide having both RNA and DNA bases.
摘要翻译：本发明涉及编码可用于促进真核细胞中的同源重组的重组酶的基因。该申请教导了可以使用一种重组酶来分离不同物种的同源重组酶的方法，以及鉴定分离的同系物的方法。来自Ustilago maydis和Saccharomyces cerevisiae的重组酶特异性地包括在本发明中。本发明包括在原核细胞中产生分离的重组酶蛋白并以活性形式回收产物的方法。本发明还包括编码非天然存在的重组酶的遗传工程改造的基因，其导致比天然存在的重组酶更高的重组速率。本发明还包括使用重组酶蛋白和重组酶基因来促进同源重组，包括宿主细胞基因组和嵌合寡核苷酸之间的重组，即具有RNA和DNA碱基的寡核苷酸。

3. 发明申请

WO1997021800A2 RECOMBINANT HELIX MODIFICATION RECOGNITION PROTEINS AND USES THEREOF 审中-公开
标题翻译：重组HELIX修饰鉴定蛋白及其用途
公开(公告)号：WO1997021800A2
公开(公告)日：1997-06-19
申请号：PCT/US1996019018
申请日：1996-11-26
申请人： THOMAS JEFFERSON UNIVERSITY , CORNELL RESEARCH FOUNDATION, INC.
发明人： THOMAS JEFFERSON UNIVERSITY , CORNELL RESEARCH FOUNDATION, INC. , KMIEC, Eric, B. , HOLLOMAN, William, K. , GERHOLD, David
IPC分类号： C12N00/00
CPC分类号： C12Q1/6827 , C07K14/47
摘要： Disclosed are the Hmp class of polypeptides, DNA sequences encoding those polypeptides, and uses thereof, particularly in methods and kits for mismatch (for example, mutation) detection.
摘要翻译：公开了多肽的Hmp类，编码那些多肽的DNA序列及其用途，特别是用于错配（例如突变）检测的方法和试剂盒。

4. 发明申请

WO1997048714A1 CHIMERIC MUTATIONAL VECTORS HAVING NON-NATURAL NUCLEOTIDES 审中-公开
标题翻译：具有非天然核的CHIMERIC MUTATIONAL VECTORS
公开(公告)号：WO1997048714A1
公开(公告)日：1997-12-24
申请号：PCT/US1997010538
申请日：1997-06-16
申请人： THOMAS JEFFERSON UNIVERSITY
发明人： THOMAS JEFFERSON UNIVERSITY , KMIEC, Eric, B.
IPC分类号： C07H21/00
CPC分类号： C07H21/00 , C12N15/10 , C12N15/8213 , C12N15/8241 , C12N15/85 , C12N15/90 , C12N2800/108
摘要： The application concerns the design and use of small, duplex oligonucleotides and oligomers of nucleotide analogs, termed Chimeric Mutational Vectors (CMV) to effect mutation in a target gene of a eukaryotic cell by homologous recombination between the CMV and the target. The CMV comprises ribonucleotides and deoxyribonucleotides and nucleotide analogs of each (generically "nucleobases"). The application discloses that CMV contain at least two segments of at least 3 ribo-type nucleobases paired to deoxyribo-type nucleobases, which regions flank the region of the CMV that corresponds to the mutation to be introduced into the target gene. The ribo-type nucleobases should preferably be nuclease resistant i.e., other than naturally occurring ribonucleotides. The uses of CMV include gene therapy of genetic diseases and construction of transgenic plants and animals.
摘要翻译：该应用涉及小型，双链寡核苷酸和称为嵌合突变载体（CMV）的核苷酸类似物寡核苷酸和寡聚体的设计和使用，以通过CMV与靶标之间的同源重组来实现真核细胞的靶基因中的突变。 CMV包含核糖核苷酸和脱氧核糖核苷酸和各自的核苷酸类似物（通常为“核碱基”）。该申请公开了CMV含有与脱氧核糖核苷酸碱基配对的至少3个核糖核苷酸碱基的至少两个片段，CMV区域的侧翼区域与待引入靶基因的突变相对应。核糖核苷酸碱基应优选为核酸酶抗性，即天然存在的核糖核苷酸以外。 CMV的用途包括遗传疾病的基因治疗和转基因植物和动物的建设。

5. 发明申请

WO1995015972A1 COMPOUNDS AND METHODS FOR SITE-DIRECTED MUTATIONS IN EUKARYOTIC CELLS 审中-公开
标题翻译：化合物和方法用于原核细胞中的位点指导突变
公开(公告)号：WO1995015972A1
公开(公告)日：1995-06-15
申请号：PCT/US1994014181
申请日：1994-12-09
申请人： THOMAS JEFFERSON UNIVERSITY
发明人： THOMAS JEFFERSON UNIVERSITY , KMIEC, Eric, B.
IPC分类号： C07H21/02
CPC分类号： A01K67/0275 , A01K2217/05 , C07H21/00 , C07K14/82 , C12N15/80 , C12N15/907
摘要： The present invention concerns a polynucleotide having both ribonucleotides and deoxyribonucleotides in a first strand and solely deoxyribonucleotides in a second strand; wherein the strands are Watson-Crick paired and are linked by an oligonucleotide so that the polynucleotide has at most a single 3' and a single 5' end. These ends can be ligated so that the polynucleotide is a single continuous circular polymer. The polynucleotide can be used to induce specific alterations in targeted genes.
摘要翻译：本发明涉及在第一链中具有核糖核苷酸和脱氧核糖核苷酸的多核苷酸，并且仅涉及第二链中的脱氧核糖核苷酸; 其中链是Watson-Crick配对的并且通过寡核苷酸连接，使得多核苷酸具有至多单个3'和单个5'末端。可以连接这些末端，使得多核苷酸是单个连续的环状聚合物。多核苷酸可用于诱导靶基因的特异性改变。

6. 发明申请

WO1997041141A1 METHODS AND COMPOUNDS FOR CURING DISEASES CAUSED BY MUTATIONS 审中-公开
标题翻译：治疗造成疾病的方法和化合物
公开(公告)号：WO1997041141A1
公开(公告)日：1997-11-06
申请号：PCT/US1997007362
申请日：1997-05-01
申请人： THOMAS JEFFERSON UNIVERSITY
发明人： THOMAS JEFFERSON UNIVERSITY , KMIEC, Eric, B. , COLE-STRAUSS, Allyson , YOON, Kyonggeun
IPC分类号： C07H21/00
CPC分类号： C12N15/113 , A61K38/00 , C07H21/00 , C07K14/805 , C12N2310/13 , C12N2310/321 , C12N2310/53 , C12N2310/3521
摘要： The invention concerns the use of duplex oligonucleotides having both 2'-deoxyribonucleotides and ribonucleotides, wherein there is base pairing between the two types of nucleotides. The sequence of the oligonucleotide is selected so that the 3' and 5' most regions of the oligonucleotides are homologous with (identical to) the sequence of a preselected target gene of a cell. The two regions of homology embrace a region that is heterologous with the target sequence. The introduction of the oligonucleotide into the nucleus of the cell causes the alteration of the target gene such that the sequence of the altered target gene is the sequence of the heterologous region. Consequently, the oligonucleotides of the invention are termed chimeric mutation vectors (CMV). In one embodiment of the invention the target gene is a globin gene and the target cell is a hematopoietic stem cell. This embodiment can be used to correct certain hemoglobinopathies such as Sickle Cell Disease, beta -thalassemia and Gaucher Disease. The rate of correction of the globin gene is high enough so that no selection of the treated hematopoietic stem cells is required to obtain a therapeutically significant effect. In one embodiment the ribonucleotides of the CMV contain methylated 2'-O.
摘要翻译：本发明涉及使用具有2'-脱氧核糖核苷酸和核糖核苷酸的双链寡核苷酸，其中两种核苷酸之间存在碱基配对。选择寡核苷酸的序列使得寡核苷酸的3'和5'大部分区域与细胞的预选靶基因的序列同源（相同）。两个同源区域包含与靶序列异源的区域。将寡核苷酸引入细胞核导致靶基因的改变，使得改变的靶基因的序列是异源区的序列。因此，本发明的寡核苷酸称为嵌合突变载体（CMV）。在本发明的一个实施方案中，靶基因是珠蛋白基因，靶细胞是造血干细胞。该实施方案可用于校正某些血红蛋白病，例如镰状细胞病，β-地中海贫血和戈谢病。球蛋白基因的校正率足够高，因此不需要选择经处理的造血干细胞来获得治疗上显着的效果。在一个实施方案中，CMV的核糖核苷酸含有甲基化的2'-O。

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