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91. 发明授权

US09644198B2 RNase H-based assays utilizing modified RNA monomers 有权
公开(公告)号：US09644198B2
公开(公告)日：2017-05-09
申请号：US13959637
申请日：2013-08-05
申请人： Joseph Alan Walder , Mark Aaron Behlke , Scott Rose , Joesph Dobosy
发明人： Joseph Alan Walder , Mark Aaron Behlke , Scott Rose , Joesph Dobosy
IPC分类号： C12N9/96 , C12Q1/68
CPC分类号： C12N9/96 , C12Q1/6844 , C12Q1/6853 , C12Q2549/101 , C12Q2525/186 , C12Q2521/301 , C12Q2525/121
摘要： The present invention pertains to novel oligonucleotide compounds for use in various biological assays, such as nucleic acid amplification, ligation and sequencing reactions. The novel oligonucleotides comprise a ribonucleic acid domain and a blocking group at or near the 3′ end of the oligonucleotide. These compounds offer an added level of specificity previously unseen. Methods for performing nucleic acid amplification, ligation and sequencing are also provided. Additionally, kits containing the oligonucleotides are also disclosed herein.

92. 发明申请

US20160040219A1 PROBES FOR IMPROVED MELT DISCRIMINATION AND MULTIPLEXING IN NUCLEIC ACID ASSAYS 有权
标题翻译：用于改进核酸测定中的熔融分析和多重反应的探针
公开(公告)号：US20160040219A1
公开(公告)日：2016-02-11
申请号：US14823288
申请日：2015-08-11
申请人： LUMINEX CORPORATION
发明人： Scott C. JOHNSON , Nicolas ARAB , Doug WHITMAN
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6818 , C12Q1/6816 , C12Q1/6823 , C12Q2521/301 , C12Q2521/327 , C12Q2525/101 , C12Q2525/121 , C12Q2525/161 , C12Q2525/186 , C12Q2525/301 , C12Q2527/107 , C12Q2537/143 , C12Q2561/113 , C12Q2565/101
摘要： Methods and compositions for the detection and quantification of nucleic acids are provided. In certain embodiments, methods involve the use of cleavable probes that comprise a ribonucleotide position that is susceptible to endoribonuclease (e.g., RNase H) cleavage in the presence of target nucleic acid molecules. Probes of the embodiments may also comprise non-natural nucleotide linked to a reporter and/or quenching moiety.
摘要翻译：提供了用于检测和定量核酸的方法和组合物。在某些实施方案中，方法涉及使用可切割探针，其可包含在靶核酸分子存在下内切核糖核酸酶（例如RNase H）裂解的核糖核苷酸位置。实施方案的探针还可以包含与报道分子和/或猝灭部分连接的非天然核苷酸。

93. 发明申请

US20150299772A1 SINGLE-STRANDED POLYNUCLEOTIDE AMPLIFICATION METHODS 审中-公开
标题翻译：单链多核苷酸放大方法
公开(公告)号：US20150299772A1
公开(公告)日：2015-10-22
申请号：US14647012
申请日：2013-12-02
申请人： Elim Biopharmaceuticals, Inc.
发明人： Yilin ZHANG
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6806 , C12P19/34 , C12Q1/6853 , C12Q2521/327 , C12Q2525/121 , C12Q2531/101 , C12Q2531/119 , C12Q2533/101
摘要： The present invention provides amplification methods for producing a population of single stranded polynucleotides from a target polynucleotide, comprising (a) extending an RNA primer in a complex comprising (i) a DNA template comprising a sequence that is complementary to the target polynucleotide, and (ii) the RNA primer, wherein the RNA primer is hybridized to the DNA template, and (b) cleaving the RNA primer with an enzyme that cleaves RNA from an RNA/DNA hybrid such that another RNA primer hybridizes to the DNA template and repeats primer extension by strand displacement.
摘要翻译：本发明提供了用于从靶多核苷酸产生单链多核苷酸群体的扩增方法，其包括（a）在复合物中延伸RNA引物，包括（i）包含与靶多核苷酸互补的序列的DNA模板和（ ii）RNA引物，其中RNA引物与DNA模板杂交，和（b）用从RNA / DNA杂交体切割RNA的酶切割RNA引物，使得另一RNA引物与DNA模板杂交并重复引物通过股骨移位延伸。

94. 发明申请

US20150291961A1 RNA-DIRECTED DNA CLEAVAGE BY THE Cas9-crRNA COMPLEX 审中-公开
标题翻译： RNA-DIRECTED DNA由Cas9-crRNA复合物清除
公开(公告)号：US20150291961A1
公开(公告)日：2015-10-15
申请号：US14743764
申请日：2015-06-18
申请人： VILNIUS UNIVERSITY
发明人： Virginijus SIKSNYS , Giedrius GASIUNAS , Tautvydas KARVELIS
IPC分类号： C12N15/113
CPC分类号： C12N15/113 , C12N9/22 , C12N15/102 , C12N15/902 , C12N15/907 , C12N2310/12 , C12N2310/14 , C12N2310/20 , C12N2310/3513 , C12N2310/531 , C12N2320/00 , C12N2800/80 , C12P19/34 , C12Q1/6811 , C12Q2521/301 , C12Q2522/101 , C12Q2525/121 , C12Q2525/151 , C12Q2525/301
摘要： Isolation or in vitro assembly of the Cas9-crRNA complex of the Streptococcus thermophilus CRISPR3/Cas system and use for cleavage of DNA bearing a nucleotide sequence complementary to the crRNA and a proto-spacer adjacent motif. Methods for site-specific modification of a target DNA molecule using an RNA-guided DNA endonuclease comprising at least one RNA sequence and at least one of an RuvC active site motif and an HNH active site motif; for conversion of Cas9 polypeptide into a nickase cleaving one strand of double-stranded DNA by inactivating one of the active sites (RuvC or HNH) in the polypeptide by at least one pint mutation; for assembly of active polypeptide-polyribonucleotides complex in vivo or in vitro; and for re-programming a Cas9-crRNA complex specificity in vitro or using a cassette containing a single repeat-spacer-repeat unit.
摘要翻译：分离或体外装配嗜热链球菌CRISPR3 / Cas系统的Cas9-crRNA复合物，并用于裂解含有与crRNA互补的核苷酸序列的DNA和原间隔物相邻基序。使用包含至少一个RNA序列和至少一个RuvC活性位点基序和HNH活性位点基序的RNA引导的DNA内切核酸酶的靶DNA分子的位点特异性修饰的方法; 用于通过使多肽中的一个活性位点（RuvC或HNH）失活至少一个品脱突变，将Cas9多肽转化成切口酶切割一条双链DNA; 用于在体内或体外组装活性多肽 - 多核苷酸复合物; 并且用于在体外重新编程Cas9-crRNA复合物特异性或使用含有单个重复 - 间隔 - 重复单元的盒。

95. 发明申请

US20150038360A1 Method for Multiplex-Detecting Chronic Myelogenous Leukemia Gene Using Cleavable Probe 有权
标题翻译：使用可切割探针多重检测慢性骨髓性白血病基因的方法
公开(公告)号：US20150038360A1
公开(公告)日：2015-02-05
申请号：US14371255
申请日：2013-01-09
申请人： Samsung Life Public Welfare Foundation
发明人： Kwang Woo Kim , Jong Won Kim , Do Hyun Nam , Chang-Seok Ki
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6886 , C12Q1/6851 , C12Q2600/158 , C12Q2600/16 , C12Q2521/327 , C12Q2525/121 , C12Q2565/1015
摘要： One embodiment of the present invention provides a detection kit for detecting a chronic myelogenous leukemia (CML) gene expression, wherein the detection kit includes a primer set which is specifically bound to the CML gene; and a cleavable probe which is specifically bound to the inside of a CML gene amplification product which is amplified by the primer set. Another embodiment of the present invention provides a method of measuring the CML gene expression by using the detection kit according to one embodiment of the present invention. The method according to one embodiment of the present invention is used to efficiently detect a low CML gene expression for CML diagnosis and prognosis diagnosis.
摘要翻译：本发明的一个实施方案提供了用于检测慢性骨髓性白血病（CML）基因表达的检测试剂盒，其中检测试剂盒包括特异性结合CML基因的引物组; 以及与通过引物组扩增的CML基因扩增产物内部特异性结合的可切割探针。本发明的另一个实施方案提供了通过使用根据本发明的一个实施方案的检测试剂盒来测量CML基因表达的方法。根据本发明的一个实施方案的方法用于有效检测低CML基因表达以进行CML诊断和预后诊断。

96. 发明申请

US20140255925A9 MODIFIED RNASE H ENZYMES AND THEIR USES 审中-公开
标题翻译：改良型RNASE H酶及其用途
公开(公告)号：US20140255925A9
公开(公告)日：2014-09-11
申请号：US13839334
申请日：2013-03-15
申请人： Joseph Alan WALDER , Mark Aaron BEHLKE , Scott D. ROSE , Joseph DOBOSY , Susan Marie RUPP
发明人： Joseph Alan WALDER , Mark Aaron BEHLKE , Scott D. ROSE , Joseph DOBOSY , Susan Marie RUPP
IPC分类号： C12Q1/68
CPC分类号： C12N9/22 , C12N9/1252 , C12N9/93 , C12N15/1096 , C12Q1/6848 , C12Q1/686 , C12Y301/26004 , C12Q2521/107 , C12Q2521/327 , C12Q2525/121
摘要： The invention provides a provides improvements to assays that employ RNase H cleavage for biological applications related to nucleic acid amplification and detection, where the RNase H has been reversibly inactivated.
摘要翻译：本发明提供对使用RNase H切割用于与核酸扩增和检测相关的生物学应用的测定法，其中RNase H已被可逆地灭活。

97. 发明申请

US20140106433A1 RNase H-Based Assays Utilizing Modified RNA Monomers 有权
标题翻译：使用修饰的RNA单体的RNA酶H基测定法
公开(公告)号：US20140106433A1
公开(公告)日：2014-04-17
申请号：US13959637
申请日：2013-08-05
申请人： Joseph Alan Walder , Mark Aaron Behlke , Scott Rose , Joesph Dobosy
发明人： Joseph Alan Walder , Mark Aaron Behlke , Scott Rose , Joesph Dobosy
IPC分类号： C12N9/96
CPC分类号： C12N9/96 , C12Q1/6844 , C12Q1/6853 , C12Q2549/101 , C12Q2525/186 , C12Q2521/301 , C12Q2525/121
摘要： The present invention pertains to novel oligonucleotide compounds for use in various biological assays, such as nucleic acid amplification, ligation and sequencing reactions. The novel oligonucleotides comprise a ribonucleic acid domain and a blocking group at or near the 3′ end of the oligonucleotide. These compounds offer an added level of specificity previously unseen. Methods for performing nucleic acid amplification, ligation and sequencing are also provided. Additionally, kits containing the oligonucleotides are also disclosed herein.
摘要翻译：本发明涉及用于各种生物测定的新型寡核苷酸化合物，例如核酸扩增，连接和测序反应。该寡核苷酸在寡核苷酸的3'末端或其附近包含核糖核酸结构域和封闭基团。这些化合物提供了先前未见的特异性增加水平。还提供了进行核酸扩增，连接和测序的方法。此外，本文还公开了含有寡核苷酸的试剂盒。

98. 发明申请

US20140087377A1 METHOD OF IDENTIFYING NUCLEIC ACID-CONTAINING OBJECT 审中-公开
标题翻译：识别含有核酸的物体的方法
公开(公告)号：US20140087377A1
公开(公告)日：2014-03-27
申请号：US14005213
申请日：2012-03-13
申请人： Han Oh Park , Jung Young Choi , Eun Su Han , Gu Young Song , Won Seok Jang
发明人： Han Oh Park , Jung Young Choi , Eun Su Han , Gu Young Song , Won Seok Jang
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6816 , C12Q1/6818 , C12Q2521/327 , C12Q2525/121 , C12Q2563/185
摘要： The present invention relates to a method of identifying nucleic acid-containing object, more precisely a method of identifying nucleic acid-containing object which comprises the following steps: (1) preparing nucleic acid-containing object having the nucleotide sequence complementary to the nucleotide sequence of RNA-dual probe; (2) reacting the nucleic acid included in the object with the buffer containing the RNA-dual probe conjugated with a reporter and a quencher respectively and RNase; and (3) detecting fluorescence generated from the reporter. The method of identifying an object of the present invention provides labeling sensitivity 100 times as high as that of the conventional method using sequencing or labeling with fluorescent materials, takes advantages of shorter analysis time, facilitates different labeling on a variety of products according to fluorescent materials, and makes possible unlimited product administration by product group and batch in real production process by differentiating the nucleotide sequence of each oligonucleotide.
摘要翻译：本发明涉及鉴定含核酸物质的方法，更准确地说，一种鉴定含核酸物质的方法，包括以下步骤：（1）制备具有与核苷酸序列互补的核苷酸序列的含核酸物质的RNA双探针; （2）将包含在物体中的核酸与分别与报告子和猝灭剂结合的RNA双重探针的缓冲液和RNase分别反应; 和（3）检测从报告物产生的荧光。识别本发明的目的的方法提供了使用荧光材料进行测序或标记的常规方法的100倍的标签灵敏度，具有较短的分析时间的优点，便于根据荧光材料对各种产品进行不同的标记并通过区分每个寡核苷酸的核苷酸序列，在实际生产过程中使产品组和批次无限制地进行产品管理。

99. 发明申请

US20140057264A1 DETECTION OF TARGET NUCLEIC ACID SEQUENCE BY PTO CLEAVAGE AND EXTENSION-DEPENDENT CLEAVAGE 审中-公开
标题翻译：通过PTO CLEAVAGE和扩展依赖性检测检测目标核酸序列
公开(公告)号：US20140057264A1
公开(公告)日：2014-02-27
申请号：US14008096
申请日：2012-03-29
申请人： SEEGENE, INC.
发明人： John Yoon Chun , Young Jo Lee
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6823 , C12Q1/6834 , C12Q1/6853 , C12Q2521/319 , C12Q2521/327 , C12Q2525/121 , C12Q2525/155 , C12Q2525/161 , C12Q2525/186 , C12Q2537/149 , C12Q2561/101 , C12Q2565/1015 , C12Q2527/107 , C12Q2561/109
摘要： The present invention relates to the detection of a target nucleic acid sequence by a PCEC (PTO Cleavage and Extension-Dependent Cleavage) assay. The present invention is characterized by generating a cleavage site for a nucleolytic enzyme on the extended duplex of which the formation is dependent on the presence of a target nucleic acid sequence. The present investion detects the occurrence of the cleavage of the extended duplex, thereby determining the presence of the target nucleic acid sequence.
摘要翻译：本发明涉及通过PCEC（PTO切割和延伸依赖性切割）测定检测靶核酸序列。本发明的特征在于在延伸的双链体上产生核分解酶的切割位点，其形成依赖于靶核酸序列的存在。本投资检测到扩展双链体切割的发生，从而确定靶核酸序列的存在。

100. 发明授权

US08394609B2 Primers and methods for the detection and discrimination of nucleic acids 有权
公开(公告)号：US08394609B2
公开(公告)日：2013-03-12
申请号：US12629045
申请日：2009-12-01
申请人： Irina Nazarenko , Ayoub Rashtchian , Joseph Solus , Richard Pires , Marlene Darfler , Gulilat Gebeyehu , Mekbib Astatke
发明人： Irina Nazarenko , Ayoub Rashtchian , Joseph Solus , Richard Pires , Marlene Darfler , Gulilat Gebeyehu , Mekbib Astatke
IPC分类号： C12P19/34
CPC分类号： C12Q1/6876 , C12P19/34 , C12Q1/6816 , C12Q1/6827 , C12Q1/686 , C12Q1/6869 , G01N21/6486 , C12Q2525/121 , C12Q2525/161 , C12Q2563/107 , C12Q2565/107
摘要： The present invention provides novel primers and methods for the detection of specific nucleic acid sequences. The primers and methods of the invention are useful in a wide variety of molecular biology applications and are particularly useful in allele specific PCR.

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