公开(公告)号：US20060074041A1

公开(公告)日：2006-04-06

申请号：US11228705

申请日：2005-09-16

申请人： Brian Johnston ,  Sergei Kazakov ,  Kevin Kisich

发明人： Brian Johnston ,  Sergei Kazakov ,  Kevin Kisich

IPC分类号： A61K48/00 ,  C07H21/02

CPC分类号： C12N15/113 ,  A61K38/00 ,  C07H21/00 ,  C12N15/1136 ,  C12N15/1138 ,  C12N2310/111 ,  C12N2310/122 ,  C12N2310/152 ,  C12N2310/315 ,  C12N2310/3511 ,  C12N2310/53 ,  C12Q1/6813

摘要： The present invention is directed to novel nucleic acid molecules and methods for their use. More specifically, the novel nucleic acid molecules of the present invention are capable of tightly and specifically interacting with a target molecule of interest not only through standard Watson-Crick base pairing, but also through additional features which allow the antisense molecules to become topologically “locked” onto the target nucleic acid, thereby imparting improved transcription and translation inhibitory properties.

摘要翻译： 本发明涉及新的核酸分子及其使用方法。 更具体地说，本发明的新型核酸分子不仅可以通过标准的沃森 - 克里克碱基配对，而且通过允许反义分子变得拓扑“锁定”的附加特征，与目标靶分子紧密而特异地相互作用 “到靶核酸上，从而赋予改善的转录和翻译抑制性质。

公开(公告)号：US20070105108A1

公开(公告)日：2007-05-10

申请号：US10561691

申请日：2004-06-25

申请人： Sergei Kazakov ,  Anne Dallas ,  Tai-Chih Kuo ,  Brian Johnston

发明人： Sergei Kazakov ,  Anne Dallas ,  Tai-Chih Kuo ,  Brian Johnston

IPC分类号： C40B40/08 ,  C40B30/06 ,  C07H21/02 ,  C07H21/04

CPC分类号： C12N15/111 ,  C12N2310/11 ,  C12N2310/122 ,  C12N2310/3519 ,  C12N2310/53 ,  C12N2320/13

摘要： The invention provides allosterically regulatable polynucleotides capable of target-dependent circularization and topological linkage to a target nucleic acid molecule. Polynucleotides of the invention include a target binding sequence and a regulatory element which prevents circularization in the absence of the target binding. Polynucleotides may include a catalytic domain, allowing circularization to proceed via catalysis when the target binding sequence of the polynucleotide is bound to the target. Topologically linked polynucleotides may be used for detection of target molecules or to inhibit transcription or translation of the target.

摘要翻译： 本发明提供能够靶向依赖性环化和与靶核酸分子拓扑连接的变构可调节多核苷酸。 本发明的多核苷酸包括靶结合序列和在没有靶结合的情况下防止环化的调节元件。 多核苷酸可以包括催化结构域，当多核苷酸的靶结合序列与靶标结合时，允许通过催化进行环化。 拓扑结合的多核苷酸可用于检测靶分子或抑制靶的转录或翻译。

公开(公告)号：US20060051789A1

公开(公告)日：2006-03-09

申请号：US11173100

申请日：2005-07-01

申请人： Sergei Kazakov ,  Alexander Vlassov ,  Anne Dallas ,  Attila Seyhan ,  Levente Egry ,  Heini Ilves ,  Roger Kaspar ,  Brian Johnston

发明人： Sergei Kazakov ,  Alexander Vlassov ,  Anne Dallas ,  Attila Seyhan ,  Levente Egry ,  Heini Ilves ,  Roger Kaspar ,  Brian Johnston

IPC分类号： C40B40/08 ,  C12P19/34

CPC分类号： C12N15/1093 ,  C12P19/34 ,  C40B40/08 ,  C40B50/06

摘要： Methods of preparing gene-specific oligonucleotide libraries are disclosed. In one embodiment a double-stranded RNA corresponding to both sense and antisense strands of mRNA is digested by ribonuclease to produce short RNA fragments. In subsequent ligation steps, flanking oligoribonucleotides of defined sequences may be attached to the 3- and 5-ends of each fragment by RNA ligase (such as T4 RNA ligase). The products of ligation can be reverse transcribed and PCR amplified (RT-PCR) using the oligonucleotides attached to the gene-derived sequences as primer-binding sites. Various methods for incorporating libraries into expression vectors allowing expression of either siRNAs or shRNAs are also disclosed.

摘要翻译： 公开了制备基因特异性寡核苷酸文库的方法。 在一个实施方案中，对应于mRNA的有义链和反义链的双链RNA被核糖核酸酶消化以产生短RNA片段。 在随后的连接步骤中，确定序列的侧翼寡核糖核苷酸可以通过RNA连接酶（例如T4 RNA连接酶）连接到每个片段的3和5末端。 连接产物可以使用连接到基因衍生序列的寡核苷酸作为引物结合位点进行逆转录和PCR扩增（RT-PCR）。 还公开了将文库掺入允许表达siRNA或shRNA的表达载体的各种方法。