Tissue culture system for production of hepatitis C virus
    1.
    发明授权
    Tissue culture system for production of hepatitis C virus 有权
    组织培养系统用于生产丙型肝炎病毒

    公开(公告)号:US08481050B2

    公开(公告)日:2013-07-09

    申请号:US12310309

    申请日:2007-08-22

    IPC分类号: A61K48/00 A61K50/00

    摘要: A tissue culture system for production of infectious hepatitis C virus is described. In particular, the invention provides recombinant monocistronic and bicistronic genomic constructs for production of virus, including constructs for production of wild-type HCV type 2a strain JFH1 and constructs for production of chimeric viruses comprising HCV proteins from strain JFH1 and a second HCV isolate. Constructs of the invention also include a reporter gene to facilitate measurement of RNA replication and viral infectivity in cultures. The cell culture system may also include various factors that improve viral replication or infectivity. In addition, a neutralization assay using HCV grown in cell culture is described.

    摘要翻译: 描述了用于生产感染性丙型肝炎病毒的组织培养系统。 特别地,本发明提供了用于产生病毒的重组单顺反子和双顺反子基因组构建体,包括用于产生野生型HCV 2a型菌株JFH1的构建体和用于产生包含来自菌株JFH1的HCV蛋白质和第二HCV分离物的嵌合病毒的构建体。 本发明的构建体还包括有利于测量培养物中RNA复制和病毒感染性的报告基因。 细胞培养系统还可以包括改善病毒复制或感染性的各种因素。 此外,描述了使用在细胞培养物中生长的HCV的中和测定。

    Modified small interfering RNA molecules and methods of use
    2.
    发明授权
    Modified small interfering RNA molecules and methods of use 有权
    修饰的小干扰RNA分子和使用方法

    公开(公告)号:US08138161B2

    公开(公告)日:2012-03-20

    申请号:US11664008

    申请日:2005-09-30

    IPC分类号: A61K48/00 C07H21/02 C07H21/04

    摘要: The present invention provides double-stranded RNA molecules that mediate RNA interference in target cells, preferably hepatic cells. The invention also provides double-stranded RNA (dsRNA) molecules that are modified to be resistant to nuclease degradation, which inactivates a virus, and more specifically, hepatitis C virus (HCV). The invention also provides a method of using these modified RNA molecules to inactivate virus in mammalian cells and a method of making modified small interfering RNAs (siRNAs) using human Dicer. The invention provides modified RNA molecules that are modified to include a dsRNA or siRNA wherein one or more of the pyrimidines in the RNA molecule are modified to include 2′-Fluorine. The invention also provides dsRNA or siRNA in which all pyrimidines are modified to include a 2′-Fluorine. The invention provides that the 2′-Fluorine dsRNA or siRNA molecule is further modified to include a two base deoxynucleotide “TT” sequence at the 3′ end of the molecule.

    摘要翻译: 本发明提供了介导靶细胞,优选肝细胞中的RNA干扰的双链RNA分子。 本发明还提供被修饰为对核酸酶降解具有抗性的双链RNA(dsRNA)分子,其使病毒失活,更具体地,丙型肝炎病毒(HCV)。 本发明还提供了使用这些修饰的RNA分子来灭活哺乳动物细胞中的病毒的方法和使用人Dicer制备修饰的小干扰RNA(siRNA)的方法。 本发明提供经修饰以包括dsRNA或siRNA的修饰的RNA分子,其中RNA分子中的一个或多个嘧啶被修饰为包括2'-氟。 本发明还提供dsRNA或siRNA,其中所有嘧啶被修饰以包括2'-氟。 本发明提供了2'-氟dsRNA或siRNA分子被进一步修饰,以在分子的3'端包括两碱基脱氧核苷酸“TT”序列。

    Modified Small Interfering Rna Molecules and Methods of Use
    3.
    发明申请
    Modified Small Interfering Rna Molecules and Methods of Use 有权
    改良的小干扰Rna分子和使用方法

    公开(公告)号:US20080269148A1

    公开(公告)日:2008-10-30

    申请号:US11664008

    申请日:2005-09-30

    IPC分类号: A61K31/70 C07H21/00 A61P31/00

    摘要: The present invention provides double-stranded RNA molecules that mediate RNA interference in target cells, preferably hepatic cells. The invention also provides double-stranded RNA (dsRNA) molecules that are modified to be resistant to nuclease degradation, which inactivates a virus, and more specifically, hepatitis C virus (HCV). The invention also provides a method of using these modified RNA molecules to inactivate virus in mammalian cells and a method of making modified small interfering RNAs (siRNAs) using human Dicer. The invention provides modified RNA molecules that are modified to include a dsRNA or siRNA wherein one or more of the pyrimidines in the RNA molecule are modified to include 2′-Fluorine. The invention also provides dsRNA or siRNA in which all pyrimidines are modified to include a 2′-Fluorine. The invention provides that the 2′-Fluorine dsRNA or siRNA molecule is further modified to include a two base deoxynucleotide “TT” sequence at the 3′end of the molecule.

    摘要翻译: 本发明提供了介导靶细胞,优选肝细胞中的RNA干扰的双链RNA分子。 本发明还提供被修饰为对核酸酶降解具有抗性的双链RNA(dsRNA)分子,其使病毒失活,更具体地,丙型肝炎病毒(HCV)。 本发明还提供了使用这些修饰的RNA分子来灭活哺乳动物细胞中的病毒的方法和使用人Dicer制备修饰的小干扰RNA(siRNA)的方法。 本发明提供经修饰以包括dsRNA或siRNA的修饰的RNA分子,其中RNA分子中的一个或多个嘧啶被修饰为包括2'-氟。 本发明还提供dsRNA或siRNA,其中所有嘧啶被修饰以包括2'-氟。 本发明提供了2'-氟dsRNA或siRNA分子进一步修饰,以在分子的3'末端包括两碱基脱氧核苷酸“TT”序列。

    HCV NS3 protein fragments having helicase activity and improved solubility
    4.
    发明授权
    HCV NS3 protein fragments having helicase activity and improved solubility 失效
    具有解旋酶活性和改善溶解性的HCV NS3蛋白片段

    公开(公告)号:US07033805B2

    公开(公告)日:2006-04-25

    申请号:US10232643

    申请日:2002-11-25

    IPC分类号: C12N9/00 C12N9/10 C07H21/04

    摘要: The Hepatitis C Virus (HCV) NS3 protein contains amino acid motifs of a serine proteinase, a nucleotide triphosphatase (NTPase), and an RNA helicase. A carboxy fragment of the HCV NS3 protein was purified and possessed RNA helicase activity. Detections from the amino terminus resulted in the protein becoming soluble. Deletions from the carboxy terminus do not result in a loss of helicase activity until at least 50 amino acids are deleted. The helicase activity requires ATP and divalent cations such as Mg2+ and Mn2+. The helicase activity was blocked by monoclonal antibody specific to the HCV NS3 protein.

    摘要翻译: 丙型肝炎病毒(HCV)NS3蛋白含有丝氨酸蛋白酶,核苷酸三磷酸酶(NTPase)和RNA解旋酶的氨基酸基序。 HCV NS3蛋白的羧基片段被纯化并具有RNA解旋酶活性。 氨基末端的检测导致蛋白质变得可溶。 从羧基末端的缺失不会导致直到至少50个氨基酸被缺失的解旋酶活性丧失。 解旋酶活性需要ATP和二价阳离子如Mg 2+和Mn 2+。 解旋酶活性被针对HCV NS3蛋白特异性的单克隆抗体阻断。

    HCV NS3 protein fragments having helicase activity and improved solubility
    6.
    发明授权
    HCV NS3 protein fragments having helicase activity and improved solubility 失效
    具有解旋酶活性和改善溶解度的HCV NS3蛋白片段

    公开(公告)号:US06194140B1

    公开(公告)日:2001-02-27

    申请号:US08529169

    申请日:1995-09-15

    IPC分类号: C12Q100

    摘要: The Hepatitis C Virus (HCV) NS3 protein contains amino acid motifs of a serine proteinase, a nucleotide triphosphatase (NTPase), and an RNA helicase. A carboxy fragment of the HCV NS3 protein was purified and possessed RNA helicase activity. Deletions from the amino terminus resulted in the protein becoming soluble. Deletions from the carboxy terminus do not result in a loss of helicase activity until at least 50 amino acids are deleted. The helicase activity requires ATP and divalent cations such as Mg2+ and Mn2+. The helicase activity was blocked by monoclonal antibody specific to the HCV NS3 protein.

    摘要翻译: 丙型肝炎病毒(HCV)NS3蛋白含有丝氨酸蛋白酶,核苷酸三磷酸酶(NTPase)和RNA解旋酶的氨基酸基序。 HCV NS3蛋白的羧基片段被纯化并具有RNA解旋酶活性。 从氨基末端的缺失导致蛋白质变得可溶。 从羧基末端的缺失不会导致直到至少50个氨基酸被缺失的解旋酶活性丧失。 解旋酶活性需要ATP和二价阳离子如Mg2 +和Mn2 +。 解旋酶活性被HCV NS3蛋白特异性的单克隆抗体阻断。