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    Mutated and bacteriophage T4 nanoparticle arrayed F1-V immunogens from Yersinia pestis as next generation plague vaccines
    5.
    发明授权
    Mutated and bacteriophage T4 nanoparticle arrayed F1-V immunogens from Yersinia pestis as next generation plague vaccines 有权
    突变和噬菌体T4纳米颗粒从鼠疫耶尔森氏菌中排列F1-V免疫原作为下一代瘟疫疫苗

    公开(公告)号：US09328149B2

    公开(公告)日：2016-05-03

    申请号：US14320731

    申请日：2014-07-01

    申请人： The Catholic University of America

    发明人： Venigalla B. Rao ,  Pan Tao

    IPC分类号： A61K39/00 ,  C07K14/24 ,  A61K39/02 ,  C07K14/005

    CPC分类号： C07K14/24 ,  A61K39/0291 ,  A61K2039/5256 ,  C07K14/005 ,  C07K2319/21 ,  C07K2319/40 ,  C07K2319/735 ,  C12N2795/10123 ,  Y02A50/407

    摘要： Techniques from two basic approaches, structure-based immunogen design and phage T4 nanoparticle delivery, are developed to construct new plague vaccines. The NH2-terminal β-strand of F1 of Yersinia pestis is transplanted to the COOH-terminus of F1 of Yersinia pestis and the NH2-terminus sequence flanking the β-strand of F1 of Yersinia pestis is duplicated to eliminate polymerization but to retain the T cell epitopes. The mutated F1 is fused to the V antigen of Yersinia pestis to thereby form a fusion protein F1mut-V mutant, which produces a completely soluble monomer. The fusion protein F1mut-V is then arrayed on phage T4 nanoparticles via a small outer capsid protein, Soc, from a T4 phage or a T4-related phage. Both the soluble and T4 decorated F1mut-V provided approximately 100% protection to mice and rats against pneumonic plague evoked by high doses of Yersinia pestis CO92.

    摘要翻译： 开发了两种基本方法，基于结构的免疫原设计和噬菌体T4纳米颗粒递送的技术来构建新的疫苗疫苗。 将鼠疫耶尔森氏菌的F1的NH2-末端和一串被移植到鼠疫耶尔森氏菌的F1的COOH-末端，并且重复鼠疫耶尔森氏菌的F1侧链的NH2-末端序列，以消除聚合，但保留 T细胞表位。 突变的F1融合到鼠疫耶尔森氏菌的V抗原，从而形成融合蛋白F1mut-V突变体，其产生完全可溶的单体。 然后通过T4噬菌体或与T4相关的噬菌体的小外壳衣壳蛋白Soc，将融合蛋白F1mut-V排列在噬菌体T4纳米颗粒上。 可溶性和T4装饰的F1mut-V都能够对小鼠和大鼠提供大约100％的抗高剂量鼠疫耶尔森氏菌CO92诱发的肺炎瘟疫。

    PROTEIN AND NUCLEIC ACID DELIVERY VEHICLES, COMPONENTS AND MECHANISMS THEREOF
    6.
    发明申请
    PROTEIN AND NUCLEIC ACID DELIVERY VEHICLES, COMPONENTS AND MECHANISMS THEREOF 有权
    蛋白质和核酸递送机构，其组成和机理

    公开(公告)号：US20130196416A1

    公开(公告)日：2013-08-01

    申请号：US13796263

    申请日：2013-03-12

    申请人： THE CATHOLIC UNIVERSITY OF AMERICA

    发明人： Venigalla B. Rao

    IPC分类号： C12N15/88

    CPC分类号： C12N15/86 ,  A61K31/7088 ,  A61K31/711 ,  A61K47/48776 ,  A61K47/6901 ,  A61K48/00 ,  C07K14/005 ,  C12N7/00 ,  C12N15/87 ,  C12N15/88 ,  C12N2795/10042 ,  C12N2795/10122 ,  C12N2795/10142 ,  C12N2795/10152

    摘要： Complex viruses are assembled from simple protein subunits by sequential and irreversible assembly. During genome packaging in bacteriophages, a powerful molecular motor assembles at the special portal vertex of an empty prohead to initiate packaging. An aspect of the invention relates to the phage T4 packaging machine being highly promiscuous, translocating DNA into finished phage heads as well as into proheads. Single motors can force exogenous DNA into phage heads at the same rate as into proheads and phage heads undergo repeated initiations, packaging multiple DNA molecules into the same head. This shows that the phage DNA packaging machine has unusual conformational plasticity, powering DNA into an apparently passive capsid receptacle, including the highly stable virus shell, until it is full. These features allow for the design of a novel class of nanocapsid delivery vehicles.

    摘要翻译： 复杂病毒通过顺序和不可逆组装由简单蛋白质亚基组装。 在噬菌体的基因组包装过程中，一个强大的分子马达组装在一个空头颅的特殊门户顶点，开始包装。 本发明的一个方面涉及高度混杂的噬菌体T4包装机，将DNA转运到成品噬菌体头以及pro头中。 单个电机可以将外源DNA以与头孢子相同的速率强制进入噬菌体头，并且噬菌体头经历反复的引发，将多个DNA分子包装在相同的头中。 这表明噬菌体DNA包装机具有不寻常的构象可塑性，将DNA提供到明显被动的衣壳容器中，包括高度稳定的病毒壳，直至其充满。 这些特征允许设计一种新型的纳米框输送车辆。

    ENGINEERING OF BACTERIOPHAGES BY GENOME EDITING USING THE CRISPR-CAS9 SYSTEM
    7.
    发明申请
    ENGINEERING OF BACTERIOPHAGES BY GENOME EDITING USING THE CRISPR-CAS9 SYSTEM 审中-公开

    公开(公告)号：US20190330643A1

    公开(公告)日：2019-10-31

    申请号：US16355932

    申请日：2019-03-18

    申请人： The Catholic University of America

    发明人： Venigalla B. Rao ,  Pan Tao

    IPC分类号： C12N15/70 ,  C12N15/11 ,  C12N9/22

    摘要： Embodiments of the invention provide systems, methods, and kits for CRISPR-based editing of DNA targets by a CRISPR-associated (Cas) enzyme. The systems include a bacterial host cell adapted to produce an engineered bacteriophage comprising a Cas protein and guide RNA that do not naturally occur together, i.e. they are engineered to occur together, as well as a DNA repair template comprising a donor DNA having a desired mutation. The guide RNA comprises a trans-activating crRNA and a guide sequence complementary to a target protospacer in a bacteriophage genome. A wild-type bacteriophage or a glucosylhydroxymethyl cytosine (ghmC)-unmodified mutant bacteriophage may be delivered into a disclosed bacterial host cell to create recombinants of bacteriophage having the desired mutation provided by the donor DNA.