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1. 发明授权

US06936467B2 Targeted chromosomal genomic alterations with modified single stranded oligonucleotides 失效
标题翻译：用修饰的单链寡核苷酸靶向染色体基因组改变
公开(公告)号：US06936467B2
公开(公告)日：2005-08-30
申请号：US09818875
申请日：2001-03-27
申请人： Eric B. Kmiec , Howard B. Gamper , Michael C. Rice
发明人： Eric B. Kmiec , Howard B. Gamper , Michael C. Rice
IPC分类号： A61K38/00 , A61K48/00 , C12N15/10 , C12N15/113 , C12N15/82 , C12N15/09
CPC分类号： C12N15/113 , A61K38/00 , A61K48/00 , C12N15/102 , C12N15/8213 , C12N2310/315 , C12N2310/321 , C12N2310/3231 , C12N2310/346 , Y02A50/473 , C12N2310/3521
摘要： Presented are methods and compositions for targeted chromosomal genomic alterations using modified single-stranded oligonucleotides. The oligonucleotides of the invention have at least one modified nuclease-resistant terminal region comprising phosphorothioate linkages, LNA analogs or 2′-O-Me base analogs.
摘要翻译：提出了使用修饰的单链寡核苷酸的靶向染色体基因组改变的方法和组合物。本发明的寡核苷酸具有包含硫代磷酸酯键，LNA类似物或2'-O-Me碱基类似物的至少一个修饰的核酸酶抗性末端区域。

2. 发明授权

US07229767B2 Genomics applications for modified OLIGO nucleotides 失效
标题翻译：用于修饰的OLIGO核苷酸的基因组学应用
公开(公告)号：US07229767B2
公开(公告)日：2007-06-12
申请号：US10672735
申请日：2003-09-26
申请人： Eric B. Kmiec , Howard B. Gamper , Michael C. Rice , Michael G. Usher
发明人： Eric B. Kmiec , Howard B. Gamper , Michael C. Rice , Michael G. Usher
IPC分类号： C07H21/04 , C12P19/34 , C12Q1/68
CPC分类号： C12Q1/6813 , C12N15/101 , C12Q1/6816 , C12Q2525/113 , C12Q2521/507
摘要： Methods for the production and use of stable complexes of duplex nucleic acid molecules and oligonucleotides are presented. These complexes can be used for the detection and purification of a known nucleic acid target as well as the manipulation of a defined nucleic acid target sequence.
摘要翻译：提出了生产和使用双链核酸分子和寡核苷酸的稳定复合物的方法。这些配合物可以用于检测和纯化已知的核酸靶，以及操作所定义的核酸靶序列。

3. 发明授权

US06573046B1 Eukaryotic use of improved chimeric mutational vectors 有权
标题翻译：改良的嵌合突变载体的真核生物学应用
公开(公告)号：US06573046B1
公开(公告)日：2003-06-03
申请号：US09429291
申请日：1999-10-28
申请人： Eric B. Kmiec , Howard B. Gamper , Allyson D. Cole-Strauss
发明人： Eric B. Kmiec , Howard B. Gamper , Allyson D. Cole-Strauss
IPC分类号： C12Q168
CPC分类号： C12N15/102
摘要： The invention is based on the reaction of recombinagenic oligonucleotides in a cell-free system containing a cytoplasmic cell extract and a test duplex DNA on a plasmid. The reaction specifically converts a mutant kanr gene to recover the resistant phenotype in transformed MutS, RecA deficient bacteria and allows for the rapid and quantitative comparison of recombinagenic oligonucleobases. Using this system a type of Duplex Mutational Vector termed a Heteroduplex Mutational Vector, was shown to be more active in than the types of mutational vectors heretofore tested. Further improvements in activity were obtained by replacement of a tetrathymidine linker by a nuclease resistant oligonucleotide, such as tetra-2′-O-methyl-uridine, to link the two strands of the Duplex Mutational Vector and removal of the DNA-containing intervening segment. The claims concern Duplex Mutational Vectors that contain the above improvements. In an alternative embodiment the claims concern a reaction mixture containing a recombinagenic oligonucleobase, a cell-free enzyme mixture and a duplex DNA containing a target sequence. In yet an alternative embodiment, the invention concerns the use of such mixture to test improvements in recombinagenic oligonucleobases, as well as to test the effects of compounds on the activity of the cell-free enzyme mixture and also to make specific changes in the target DNA sequence.
摘要翻译：本发明基于重组型寡核苷酸在含有质粒上的细胞质细胞提取物和测试双链体DNA的无细胞系统中的反应。该反应特异性转化突变型kanr基因以在转化的MutS，RecA缺陷型细菌中恢复抗性表型，并允许对重组型寡核苷酸的快速和定量比较。使用该系统，称为异源双链突变载体的一种双链突变载体被证明比迄今为止测试的突变载体的类型更有活性。通过用核酸酶抗性寡核苷酸（例如四 - 2'-O-甲基 - 尿苷）替代四硫氨酸接头，连接双链突变载体的两条链和去除含有DNA的中间片段，获得活性的进一步改善。这些权利要求涉及包含上述改进的双链突变载体。在替代实施方案中，权利要求涉及含有重组型寡核苷酸，无细胞的酶混合物和含有靶序列的双链体DNA的反应混合物。在另一个实施方案中，本发明涉及使用这样的混合物来测试重组型寡核苷酸的改进，以及测试化合物对无细胞酶混合物的活性的影响，并且还对靶DNA进行特异性改变序列。

4. 发明授权

US6010907A Eukaryotic use of non-chimeric mutational vectors 失效
标题翻译：非嵌合突变载体的真核生物学应用
公开(公告)号：US6010907A
公开(公告)日：2000-01-04
申请号：US78064
申请日：1998-05-12
申请人： Eric B. Kmiec , Howard B. Gamper , Allyson D. Cole-Strauss
发明人： Eric B. Kmiec , Howard B. Gamper , Allyson D. Cole-Strauss
IPC分类号： C12N15/09 , C12N1/11 , C12N15/10 , C12N5/10 , C12N15/11 , C12N15/63
CPC分类号： C12N15/102
摘要： The invention is based on the reaction of Duplex Mutational Vector in a cell-free system containing a cytoplasmic cell extract and a test plasmid. The reaction specifically converts a mutant kan.sup.r gene to recover the resistant phenotype in transformed MutS, RecA deficient bacteria. Using this system a type of Duplex Mutational Vector termed a Non-Chimeric Mutational Vector, having no RNA:DNA hybrid-duplex is shown to be an effective substrate for eukaryotic enzymes. The invention concerns the use of Non-Chimeric Mutational Vectors protected from 3' exonuclease attack in eukaryotic cells. Such protection can be conferred by replacement of a tetrathymidine linker by a nuclease resistant oligonucleotide, such as tetra-2'-O-methyl-uridine, to link the two strands of the recombinagenic oligonucleobase.
摘要翻译：本发明基于双链突变载体在含有细胞质细胞提取物和测试质粒的无细胞系统中的反应。该反应特异性转化突变型kanr基因以在转化的MutS，RecA缺陷型细菌中恢复抗性表型。使用该系统，一种称为非嵌合突变载体的双链突变载体，没有RNA：DNA杂合双链体显示为真核生物酶的有效底物。本发明涉及在真核细胞中保护3'核酸外切酶攻击的非嵌合突变载体的用途。这样的保护可以通过用核酸酶抗性寡核苷酸（例如四 - 2'-O-甲基 - 尿苷）替代四连体酶连接体来连接重组型寡核苷酸的两条链。

5. 发明授权

US07112405B2 Compositions and methods for enhancing oligonucleotide-mediated gene alteration 失效
公开(公告)号：US07112405B2
公开(公告)日：2006-09-26
申请号：US10260375
申请日：2002-09-27
申请人： Eric B. Kmiec , Michael C. Rice , Li Liu
发明人： Eric B. Kmiec , Michael C. Rice , Li Liu
IPC分类号： C12Q1/68 , G01N33/53
CPC分类号： C12N15/102 , A61K38/00
摘要： Composition and methods for enhancing oligonucleotide-directed nucleic acid sequence alteration in vivo, ex vivo and in vitro are presented. These methods and compositions involve cells and cell-free extracts with altered levels or activities of a protein from the RAD52 epistasis group, the mismatch repair group and/or the excision repair group.

6. 发明授权

US06210916B1 Rec 2 kinase 失效
标题翻译： Rec 2激酶
公开(公告)号：US06210916B1
公开(公告)日：2001-04-03
申请号：US09587436
申请日：2000-06-05
申请人： Pamela A. Havre , Michael C. Rice , William K. Holloman , Eric B. Kmiec
发明人： Pamela A. Havre , Michael C. Rice , William K. Holloman , Eric B. Kmiec
IPC分类号： C12Q148
CPC分类号： C12N9/1205 , C12N2799/026 , C12Q1/48
摘要： The invention includes a method of phosphorylating a serine containing substrate by incubating the substrate with ATP and an enzyme that is hsRec2 or muRec2 or a derivative thereof. The natural substrates of the kinase activity of Rec2 are the cell cycle control proteins such as p53 and cyclin E. The over expression of Rec2 is known to cause cell-cycle arrest and apoptosis and the invention discloses that these effects are kinase mediated. Accordingly, the invention provides a method of assessing antagonists and agonists of Rec2, which antagonists and agonists would have pharmacological activity. The invention further discloses that there is specific binding between hsRec2 and at least three cell cycle control proteins: p53, PCNA and cdc2.
摘要翻译：本发明包括通过用ATP孵育底物和使用hsRec2或muRec2或其衍生物的酶来磷酸化含丝氨酸的底物的方法。 Rec2的激酶活性的天然底物是细胞周期控制蛋白如p53和细胞周期蛋白E.已知Rec2的过度表达引起细胞周期阻滞和凋亡，本发明公开了这些作用是激酶介导的。因此，本发明提供了一种评估Rec2的拮抗剂和激动剂的方法，该拮抗剂和激动剂将具有药理活性。本发明还公开了hsRec2与至少三种细胞周期控制蛋白之间的特异性结合：p53，PCNA和cdc2。

7. 发明授权

US06410226B1 Mammalian and human REC2 失效
标题翻译：哺乳动物和人类REC2
公开(公告)号：US06410226B1
公开(公告)日：2002-06-25
申请号：US08927165
申请日：1997-09-11
申请人： Eric B. Kmiec , William K. Holloman , Michael C. Rice , Sheryl T. Smith , Zhigang Shu
发明人： Eric B. Kmiec , William K. Holloman , Michael C. Rice , Sheryl T. Smith , Zhigang Shu
IPC分类号： C12Q168
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 irradiation 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 thymidien 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的过度表达促进同源重组，特别是使用DNA / RNA嵌合寡核苷酸的同源重组，并使细胞对照射的凋亡作用敏感。发现REC2启动子与强增强子例如SV40增强子组合，在细胞照射后是强启动子。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腺嘧啶激酶基因的基因来使细胞对辐射治疗敏感。

8. 发明授权

US06174694B1 REC2 kinase 失效
标题翻译： REC2激酶
公开(公告)号：US06174694B1
公开(公告)日：2001-01-16
申请号：US09157603
申请日：1998-09-21
申请人： Pamela A. Havre , Michael C. Rice , William K. Holloman , Eric B. Kmiec
发明人： Pamela A. Havre , Michael C. Rice , William K. Holloman , Eric B. Kmiec
IPC分类号： C12Q148
CPC分类号： C12N9/1205 , C12N2799/026 , C12Q1/48
摘要： The invention includes a method of phosphorylating a serine containing substrate by incubating the substrate with ATP and an enzyme that is hsRec2 or muRec2 or a derivative thereof. The natural substrates of the kinase activity of Rec2 are the cell cycle control proteins such as p53 and cyclin E. The over expression of Rec2 is known to cause cell-cycle arrest and apoptosis and the invention discloses that these effects are kinase mediated. Accordingly, the invention provides a method of assessing antagonists and agonists of Rec2, which antagonists and agonists would have pharmacological activity. The invention further discloses that there is specific binding between hsRec2 and at least three cell cycle control proteins: p53, PCNA and cdc2.
摘要翻译：本发明包括通过用ATP孵育底物和使用hsRec2或muRec2或其衍生物的酶来磷酸化含丝氨酸的底物的方法。 Rec2的激酶活性的天然底物是细胞周期控制蛋白如p53和细胞周期蛋白E.已知Rec2的过度表达引起细胞周期阻滞和凋亡，本发明公开了这些作用是激酶介导的。因此，本发明提供了一种评估Rec2的拮抗剂和激动剂的方法，该拮抗剂和激动剂将具有药理活性。本发明还公开了hsRec2与至少三种细胞周期控制蛋白之间的特异性结合：p53，PCNA和cdc2。

9. 发明授权

US07468244B2 Polymorphism detection and separation 失效
标题翻译：多态性检测和分离
公开(公告)号：US07468244B2
公开(公告)日：2008-12-23
申请号：US10260150
申请日：2002-09-27
申请人： Eric B. Kmiec , Michael C. Rice
发明人： Eric B. Kmiec , Michael C. Rice
IPC分类号： C07H21/04 , C12P19/34 , C12Q1/68
CPC分类号： C12Q1/6806 , C12Q2521/507 , C12Q2522/101
摘要： Methods and compositions for polymorphism detection and separation. The methods are readily multiplexed, can be adapted to a variety of existing detection systems, and permit target amplification without PCR. The methods permit allelic variants selectively to be isolated, with or without contemporaneous detection, and finds particular utility in facilitating the construction of coisogenic cell collections in which the cells differ genotypically by single nucleotide changes targeted to defined loci.
摘要翻译：用于多态性检测和分离的方法和组合物。这些方法易于复用，可以适应各种现有的检测系统，并且允许无PCR的靶扩增。该方法允许在有或没有同时检测的情况下选择性地分离等位基因变体，并且在促进构建细胞集合的特定用途中，其中细胞基因型通过靶向定义的基因座的单核苷酸变化而基因型不同。

10. 发明授权

US5888983A Method and oligonucleobase compounds for curing diseases caused by mutations 失效
标题翻译：用于治疗由突变引起的疾病的方法和寡核苷酸化合物
公开(公告)号：US5888983A
公开(公告)日：1999-03-30
申请号：US906265
申请日：1997-08-05
申请人： Eric B. Kmiec , Allyson D. Cole-Strauss
发明人： Eric B. Kmiec , Allyson D. Cole-Strauss
IPC分类号： A61K38/00 , C07H21/00 , C07K14/805 , C12N15/10 , C12N15/113 , C12N15/82 , C12N15/85 , C12N15/90 , A61K48/00 , C12N15/11 , C12Q1/68
CPC分类号： C12N15/113 , C07H21/00 , C07K14/805 , C12N15/10 , C12N15/8213 , C12N15/8241 , C12N15/85 , C12N15/90 , A61K38/00 , C12N2310/13 , C12N2310/321 , C12N2310/53 , C12N2800/108
摘要： The invention concerns methods of introducing specific alterations in the genome of cells that have been removed from a subject suffering from a medical condition that is the result of a genetic lesion. The specific alteration is designed to correct the genetic lesion. The method comprises introducing an oligonucleotide compound, containing ribonucleotides and deoxyribonucleotides, into the cells and thereafter reintroducing the cells into the subject. Specific types of cells include hematopoietic stem cells and hepatocytes. The ribonucleotides of the compound can have 2-substituents that enhance their resistance to RNase.
摘要翻译：本发明涉及在已经从患有遗传损伤的结果的医学病症的受试者中移除的细胞基因组中引入特异性改变的方法。具体改变旨在纠正遗传病变。该方法包括将含有核糖核苷酸和脱氧核糖核苷酸的寡核苷酸化合物引入细胞，然后将细胞重新引入受试者。特定类型的细胞包括造血干细胞和肝细胞。该化合物的核糖核苷酸可以具有增强其对RNA酶的抗性的2-取代基。

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