公开(公告)号：US07883702B2

公开(公告)日：2011-02-08

申请号：US11661134

申请日：2005-08-25

申请人： Renu Wadhwa ,  Kazunari Taira ,  Sunil Kaul

发明人： Renu Wadhwa ,  Kazunari Taira ,  Sunil Kaul

IPC分类号： A61K39/395 ,  C07K16/30 ,  C07K16/46

CPC分类号： A61K31/713 ,  A61K31/7088 ,  A61K31/7105 ,  A61K45/06 ,  A61K2039/505 ,  C07K16/18 ,  C07K2317/77 ,  C12Q1/6869 ,  G01N33/5011 ,  G01N33/57484 ,  C12Q2565/607

摘要： The present invention relates to cancer therapies using an antibody that binds to mortalin 2 and a functional nucleic acid. Mortalin expression was found to be upregulated in immortalized cells and tumor tissues. Immortalized human cells highly expressing mortalin showed anchorage-independent growth. When the K antibody, which is a specific anti-mortalin antibody, was injected into a tumor of a nude mouse, tumor growth was suppressed or the tumor shrank compared with the case of a control. In accordance with the present invention, the use of a specific anti-mortalin antibody (K antibody) for tumor therapies and the use of such antibody as a carrier molecule for transportation of immunotoxicin and the like into cells are provided. It has been shown that mortalin can be a target for cancer therapies. In accordance with the present invention, a novel and effective anticancer agent is provided. In addition, an anti-mortalin antibody that is internalized by cells is developed. Thus, various applications using such antibody are provided.

摘要翻译： 本发明涉及使用结合赖mort啉2的抗体和功能性核酸的癌症疗法。 在永生化细胞和肿瘤组织中，发现莫达林表达上调。 高度表达唾液霉素的永生化人细胞显示出与锚固无关的生长。 当将特异性抗赖mort啉蛋白抗体的K抗体注射到裸小鼠的肿瘤中时，与对照的情况相比，抑制肿瘤生长或肿瘤缩小。 根据本发明，提供了特异性抗赖mort啉蛋白抗体（K抗体）用于肿瘤治疗的用途，以及使用这种抗体作为载体分子将免疫毒素等运送到细胞中。 已经表明，凡士林可以成为癌症治疗的靶标。 根据本发明，提供了一种新颖有效的抗癌剂。 此外，开发了由细胞内化的抗赖mort啉蛋白抗体。 因此，提供了使用这种抗体的各种应用。

公开(公告)号：US20060128617A1

公开(公告)日：2006-06-15

申请号：US10543078

申请日：2004-01-23

申请人： Michinori Kohara ,  Tsunamasa Watanabe ,  Kazunari Taira ,  Makoto Miyagishi ,  Masayuki Sudo

发明人： Michinori Kohara ,  Tsunamasa Watanabe ,  Kazunari Taira ,  Makoto Miyagishi ,  Masayuki Sudo

IPC分类号： A61K38/16 ,  A61K48/00 ,  C07H21/02 ,  C12Q1/70 ,  C12N15/86 ,  C07K14/18

CPC分类号： C12N15/1131 ,  A61K31/7105 ,  A61K38/00 ,  C12N2310/3181

摘要： A method of inhibiting the replication ability of a hepatitis C virus (HCV) is provided. An oligoribonucleotide or a peptide nucleic acid which sequence-specifically binds to the HCV-RNA, and a therapeutic agent for hepatitis C which contains any of these components as an active ingredient are provided.

摘要翻译： 提供了抑制丙型肝炎病毒（HCV）的复制能力的方法。 提供了序列特异性结合HCV-RNA的寡核糖核苷酸或肽核酸，以及含有任何这些成分作为活性成分的丙型肝炎治疗剂。

公开(公告)号：US20050197315A1

公开(公告)日：2005-09-08

申请号：US11128161

申请日：2005-05-13

申请人： Kazunari Taira ,  Makoto Miyagishi

发明人： Kazunari Taira ,  Makoto Miyagishi

IPC分类号： C12N15/11 ,  A61K48/00 ,  C07H21/02 ,  C12Q1/68

CPC分类号： C12N15/111 ,  A01K2217/05 ,  C12N2310/111 ,  C12N2310/14 ,  C12N2310/53 ,  C12N2330/30 ,  C12N2330/31

摘要： The in vivo siRNA expression system according to this invention is a system that intracellularly expresses small interfering (si) RNAs and comprises antisense and sense code DNAs coding for antisense and sense RNAs targeting any region of a target gene mRNA and one or more promoters that function to express the antisense and sense RNAs from the antisense and sense code DNAS, respectively.

摘要翻译： 根据本发明的体内siRNA表达系统是细胞内表达小干扰（si）RNA并且包含编码靶向靶基因mRNA的任何区域的反义和正义RNA的反义和有义编码DNA的系统，以及一种或多种起作用的启动子 分别从反义和感觉代码DNAS表达反义和有义RNA。

公开(公告)号：US20050053583A1

公开(公告)日：2005-03-10

申请号：US10926011

申请日：2004-08-26

申请人： Suehiro Sakaguchi ,  Kazunari Taira

发明人： Suehiro Sakaguchi ,  Kazunari Taira

IPC分类号： C12N15/09 ,  A01K67/027 ,  A61K31/7105 ,  A61K38/00 ,  A61K45/00 ,  A61K48/00 ,  A61P25/28 ,  A61P43/00 ,  C12N15/113

CPC分类号： C12N15/113 ,  A61K38/00 ,  C12N2310/111 ,  C12N2310/14 ,  C12N2799/021

摘要： By inhibiting expression of a normal prion protein in prion-infected cells by using siRNA, it is possible to inhibit accumulation of an abnormal prion protein consequently. The inhibition of the abnormal prion protein accumulation can be applied in prevention and treatment of the prion diseases. This provides a method of inhibiting the abnormal prion protein accumulation. This method is applicable in preventing and treating the prion diseases whose effective method of treating has not been established at this moment. Further, use of this method is provided herein.

摘要翻译： 通过使用siRNA抑制朊病毒感染细胞中正常朊病毒蛋白的表达，可以抑制异常朊病毒蛋白的积聚。 异常朊病毒蛋白积聚的抑制可用于朊病毒疾病的预防和治疗。 这提供了抑制异常朊蛋白积累的方法。 该方法适用于预防和治疗目前尚未建立有效治疗方法的朊病毒疾病。 此外，本文提供了该方法的使用。

公开(公告)号：US5843658A

公开(公告)日：1998-12-01

申请号：US612069

申请日：1996-03-07

申请人： Hisatoshi Uchiyama ,  Masaki Jibu ,  Kenichi Hirano ,  Kazunari Taira

发明人： Hisatoshi Uchiyama ,  Masaki Jibu ,  Kenichi Hirano ,  Kazunari Taira

IPC分类号： C07H21/00 ,  C12Q1/68 ,  C12P19/34

CPC分类号： C07H21/00 ,  C12Q1/6818

摘要： The present invention provides an in situ monitoring of the decomposing activity against oligonucleotides target in a biological tissue. A single-chain oligonucleotide target comprises an appropriate number of nucleic acid bases to be examined with an energy donor and an energy acceptor respectively at its 5'- and 3'-terminals. Monitoring of the fluorescence changes of the target after injection into a biological tissue, particularly the fluorescent resonance energy transfer(FRET) phenomena between the energy donor and acceptor, indicates whether the oligonucleotide is not decomposed yet.

摘要翻译： 本发明提供了对生物组织中针对寡核苷酸靶的分解活性的原位监测。 单链寡核苷酸靶包含在其5'和3'末端分别与能量供体和能量受体一起检查的合适数量的核酸碱基。 监测注射到生物组织中的靶的荧光变化，特别是能量供体和受体之间的荧光共振能量转移（FRET）现象，指示寡核苷酸是否尚未分解。

公开(公告)号：US5500357A

公开(公告)日：1996-03-19

申请号：US55390

申请日：1993-05-03

申请人： Kazunari Taira ,  Satoshi Nishikawa ,  Hidekatsu Maeda

发明人： Kazunari Taira ,  Satoshi Nishikawa ,  Hidekatsu Maeda

IPC分类号： C12N15/113 ,  C12N9/22 ,  C12N15/74 ,  C12N15/82 ,  C12N15/85

CPC分类号： C12N15/113 ,  C12N2310/111 ,  C12N2310/12 ,  C12N2310/121 ,  C12N2310/127

摘要： The invention provides a recombinant plasmid containing a sequence encoding any genes inserted between 5' and 3' self-cleavage ribozymes. The recombinant plasmid can be amplified in vivo as well as in vitro while growing the host cell. When obtaining RNA transcripts of the inserted sequence, the recombinant plasmid does not require a restriction enzyme digestion step (run-off transcription) since cis-acting ribozymes perform self-catalyzed cleavage at 5' and 3' sides of the inserted sequence once it is transcribed. In this specific example, the trans-acting RNA enzyme sequence is inserted between 5' and 3' cleavage ribozymes. However, the trans-acting ribozyme sequence in the recombinant plasmid can be replaceable with any other sequence (e.g., antisense RNA, RNAs of HIV-1, HDV and other RNA viruses etc.). This construct is especially useful since each unit, consisting of 5' processing ribozyme, inserted sequence, and 3' processing ribozyme, can be connected in tandem. By so doing, ribozymes targeted to various sites can initially be transcribed as a long RNA chain which subsequently undergoes cleavage to produce independent trans-acting ribozymes, each possessing a specific target site.

摘要翻译： 本发明提供了含有编码插入5'和3'自切割核酶之间的任何基因的序列的重组质粒。 重组质粒可以在生长宿主细胞的同时体内和体外扩增。 当获得插入序列的RNA转录物时，重组质粒不需要限制酶消化步骤（径流转录），因为顺式作用核酶在插入序列的5'和3'侧进行自催化裂解，一旦它是 转录。 在该具体实例中，将反式RNA酶序列插入5'和3'切割核酶之间。 然而，重组质粒中的反式核酶序列可以用任何其它序列（例如，反义RNA，HIV-1，HDV和其他RNA病毒等的RNA）来替代。 这种构建是特别有用的，因为由5'加工核酶，插入序列和3'加工核酶组成的每个单元可以串联连接。 通过这样做，靶向各种位点的核酶最初可以被转录为长的RNA链，其随后经历切割以产生独立的反式作用核酶，每个具有特定的靶位点。

公开(公告)号：US20060088837A1

公开(公告)日：2006-04-27

申请号：US10512386

申请日：2003-04-25

申请人： Kazunari Taira ,  Hiroaki Kawasaki

发明人： Kazunari Taira ,  Hiroaki Kawasaki

IPC分类号： C12Q1/68 ,  A01K67/027 ,  C07H21/02 ,  A61K48/00

CPC分类号： A01K67/0275 ,  A01K2207/15 ,  A01K2217/00 ,  A01K2217/05 ,  A01K2217/075 ,  A01K2217/20 ,  A01K2227/101 ,  A01K2227/103 ,  A01K2227/105 ,  A01K2227/106 ,  A01K2227/107 ,  A01K2227/108 ,  A61K31/7088 ,  A61K48/00 ,  A61K2121/00 ,  C12N15/111 ,  C12N15/85 ,  C12N2310/111 ,  C12N2310/14 ,  C12N2310/3519 ,  C12N2310/53 ,  C12N2330/30 ,  C12N2830/00 ,  C12N2830/003 ,  C12N2830/85

摘要： The present inventors discovered that an RNA molecule can be efficiently transferred into cytoplasm and exert RNAi effects, by producing a stem loop RNA molecule from the DNA that encodes this RNA molecule, with the use of a tRNA promoter. Also, the RNAi effects can be exerted effectively, by introducing a cytoplasm translocation signal sequence into the DNA that encodes a stem loop RNA molecule. Moreover, the RNAi effects can be exerted effectively by transferring a transcriptional product into the cytoplasm, using a pol II-type promoter. In this case, cytotoxicity can be reduced by co-expressing a Dicer gene. Furthermore, an effective dsRNA can be constructed by treating a dsRNA or a stem loop RNA molecule with Dicer protein. Knockout cells lacking a gene of interest can be conveniently constructed, using these stem-loop RNA molecule expression systems.

摘要翻译： 本发明人通过使用tRNA启动子，通过从编码该RNA分子的DNA产生茎环RNA分子，可以有效地将RNA分子转移到细胞质中并发挥RNAi作用。 此外，通过将细胞质易位信号序列引入到编码茎环RNA分子的DNA中，可以有效地发挥RNAi作用。 此外，通过使用pol II型启动子将转录产物转移到细胞质中可以有效地发挥RNAi作用。 在这种情况下，可以通过共表达Dicer基因来减少细胞毒性。 此外，通过用Dicer蛋白处理dsRNA或茎环RNA分子可以构建有效的dsRNA。 使用这些茎环RNA分子表达系统，可以方便地构建缺乏目的基因的敲除细胞。

公开(公告)号：US06524853B1

公开(公告)日：2003-02-25

申请号：US09720201

申请日：2000-12-22

申请人： Michinori Kohara ,  Kyoko Kohara ,  Kazunari Taira ,  Junichi Matsuzaki ,  Hiroshi Ohmori

发明人： Michinori Kohara ,  Kyoko Kohara ,  Kazunari Taira ,  Junichi Matsuzaki ,  Hiroshi Ohmori

IPC分类号： C12N500

CPC分类号： C07K14/005 ,  C12N7/00 ,  C12N15/1096 ,  C12N15/66 ,  C12N2770/24221 ,  C12N2770/24222

摘要： This invention provides a vector comprising cDNA encoding an RNA viral gene, characterized in that the vector is constructed such that both termini of the RNA viral gene can be transcribed precisely and uniformly. This vector is useful to elucidate the mechanisms of RNA virus replication and of the development of RNA virus infections, and to develop therapeutic agents and means of treatment, and so on.

摘要翻译： 本发明提供了包含编码RNA病毒基因的cDNA的载体，其特征在于构建载体，使RNA病毒基因的两个末端能够精确而均匀地转录。 该载体可用于阐明RNA病毒复制和RNA病毒感染发展的机制，并开发治疗剂和治疗手段等。

公开(公告)号：US20060247193A1

公开(公告)日：2006-11-02

申请号：US10544761

申请日：2004-02-10

申请人： Kazunari Taira ,  Hiroaki Kawasaki

发明人： Kazunari Taira ,  Hiroaki Kawasaki

IPC分类号： A61K48/00 ,  C12N15/09

CPC分类号： C12N15/113 ,  C12N2310/14 ,  C12N2310/141 ,  C12N2310/531

摘要： The present invention provides products and methods for modulating expression of a target gene in a cell. One such method includes introducing into the cell a polynucleotide that forms a duplex region with an mRNA transcribed from said target gene, where the duplex region comprises a mammalian miRNA target region. Another such method includes introducing into the cell an siRNA that forms a duplex region with an miRNA, or precursor thereof, where an mRNA transcribed from the target gene comprises a miRNA target region. In certain preferred embodiments, the methods further include measuring expression of the target gene. The methods are particularly useful for modulating ontogenesis, function, differentiation and/or viability of a mammalian cell. As such, the invention also provides methods for controlling ontogenesis of mammal, function of mammalian cell, differentiation of mammalian cell or viability of mammalian cell in the post-transcriptional phase by introducing into the cell a miRNA or a siRNA silencing precursor to the miRNA. The invention additionally provides polynucleotides, including vectors, useful in the method of the instant invention. The provided polynucleotides include a plasmid vector comprising a promoter and a polynucleotide sequence expressing miRNA or precursor to the miRNA. Also included is a plasmid vector comprising a promoter and a nucleotide sequence expressing siRNA silencing precursor to miRNA. In certain preferred embodiments, the mRNA is capable of forming a duplex region with an mRNA transcribed from a mammalian target gene.

摘要翻译： 本发明提供调节细胞中靶基因表达的产物和方法。 一种这样的方法包括向细胞中引入与从所述靶基因转录的mRNA形成双链体区的多核苷酸，其中双链区包含哺乳动物miRNA靶区。 另一种这样的方法包括向细胞中引入与miRNA或其前体形成双链体区的siRNA，其中从靶基因转录的mRNA包含miRNA靶区。 在某些优选实施方案中，所述方法还包括测量靶基因的表达。 该方法对于调节哺乳动物细胞的发生，功能，分化和/或活力特别有用。 因此，本发明还提供了通过将miRNA或siRNA沉默前体引入细胞来控制哺乳动物细胞的发生，哺乳动物细胞的功能，哺乳动物细胞的分化或哺乳动物细胞在转录后阶段的存活力的方法。 本发明另外提供了可用于本发明方法的多核苷酸，包括载体。 所提供的多核苷酸包括质粒载体，其包含启动子和表达miRNA或miRNA前体的多核苷酸序列。 还包括质粒载体，其包含启动子和表达miRNA沉默前体的核苷酸序列。 在某些优选的实施方案中，mRNA能够与从哺乳动物靶基因转录的mRNA形成双链体区域。

公开(公告)号：US20050191626A1

公开(公告)日：2005-09-01

申请号：US10477373

申请日：2002-05-10

申请人： Kazunari Taira ,  Satoshi Fujita ,  Shinya Yoshizaki ,  Masaki Warashina

发明人： Kazunari Taira ,  Satoshi Fujita ,  Shinya Yoshizaki ,  Masaki Warashina

IPC分类号： C12N15/10 ,  G01N33/68 ,  C12Q1/68 ,  C07K14/00 ,  C12N15/09

CPC分类号： G01N33/68 ,  C12N15/1041 ,  C12N15/1048 ,  C12N15/1062 ,  C12N15/1075 ,  C12Q2522/101 ,  C12Q2525/179 ,  C12Q2525/205 ,  C12Q2563/131

摘要： A stable linkage between a genotype and a phenotype in a cell-free system was successfully achieved by using interaction between a RNA-binding protein and RNA, between a DNA-binding protein and DNA, or by using a protein that inactivates a ribosome. Furthermore, it was found that functional proteins could be selected by using these stable linkages.

摘要翻译： 通过使用RNA结合蛋白和RNA之间，DNA结合蛋白和DNA之间的相互作用，或通过使用使核糖体失活的蛋白质，能够成功地实现无细胞系统中基因型与表型之间的稳定连锁。 此外，发现可以通过使用这些稳定连接来选择功能性蛋白质。