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    Compositions and methods for transient expression of recombinant RNA
    1.
    发明授权
    Compositions and methods for transient expression of recombinant RNA 有权

    公开(公告)号：US09951349B2

    公开(公告)日：2018-04-24

    申请号：US14343817

    申请日：2012-09-27

    申请人： Yale University

    发明人： Peter M. Rabinovich ,  Sherman M. Weissman

    IPC分类号： C12N15/85 ,  C12N15/86

    CPC分类号： C12N15/85 ,  C12N15/86 ,  C12N2760/18843 ,  C12N2760/18852

    摘要： Compositions for transient but prolonged exogenous mRNA expression through the use of the transcription system of negative strand RNA viruses, and methods of use thereof are disclosed. In some embodiments, the system contains only RNAs and does not include any DNA molecules. The compositions typically include an RNA template unit (rTeUn) that includes a virus regulatory sequences operably linked to a coding sequence of interest. The rTeUn is typically transfected to a host cell's cytoplasm in the presence of virus expression system proteins that mediate replication of the rTeUn and transcription of the transgene. The rTeUn RNA bonded to viral proteins exhibits high resistance to degradation, prolonged duration of expression, and is free of viral genes. The compositions can be used to reprogram cell. For example, the compositions and methods can be used to redirected lymphocytes to target cancer cells, or to dedifferentiate somatic cells into induce pluripotent stem cells.

    METHOD OF DE-DIFFERENTIATING AND RE-DIFFERENTIATING SOMATIC CELLS USING RNA
    2.
    发明申请
    METHOD OF DE-DIFFERENTIATING AND RE-DIFFERENTIATING SOMATIC CELLS USING RNA 有权
    使用RNA去除和重新分化肌细胞的方法

    公开(公告)号：US20160230188A1

    公开(公告)日：2016-08-11

    申请号：US15012285

    申请日：2016-02-01

    申请人： Yale University

    发明人： Peter M. Rabinovich ,  Sherman M. Weissman

    IPC分类号： C12N15/85 ,  A61K35/33 ,  A61K35/36 ,  A61K35/17

    CPC分类号： C12N15/85 ,  A61K35/17 ,  A61K35/33 ,  A61K35/36 ,  A61K48/005 ,  A61K2035/124 ,  C12N5/0696 ,  C12N2501/602 ,  C12N2501/603 ,  C12N2501/604 ,  C12N2501/606 ,  C12N2506/094 ,  C12N2506/1307 ,  C12N2510/00

    摘要： RNA prepared by in vitro transcription using a polymerase chain reaction (PCR)-generated template can be introduced into a cell to modulate cell activity. This method is useful in de-differentiating somatic cells to pluripotent, multipotent, or unipotent cells; re-differentiating stem cells into differentiated cells; or reprogramming of somatic cells to modulate cell activities such as metabolism. Cells can also be transfected with inhibitory RNAs, such as small interfering RNA (siRNA) or micro RNA (miRNA), or combinations thereof to induce reprogramming of somatic cells. For example, target cells are isolated from a donor, contacted with one or more RNA's causing the cells to be de-differentiated, re-differentiated, or reprogrammed in vitro, and administered to a patient in need thereof. The resulting cells are useful for treating one or more symptoms of a variety of diseases and disorders, for organ regeneration, and for restoration of the immune system.

    摘要翻译： 通过使用聚合酶链反应（PCR）生成的模板通过体外转录制备的RNA可以引入细胞以调节细胞活性。 该方法可用于将体细胞去分化为多能，多能或单能细胞; 将干细胞重新分化成分化细胞; 或重编程体细胞以调节细胞活性如代谢。 还可以用抑制性RNA如小干扰RNA（siRNA）或微RNA（miRNA）或其组合来转染细胞以诱导体细胞的重编程。 例如，从供体分离靶细胞，与一个或多个RNA接触，导致细胞在体外被去分化，再分化或重新编程，并施用于有需要的患者。 所得细胞可用于治疗各种疾病和病症的一种或多种症状，用于器官再生和用于免疫系统的恢复。

    Compositions and Methods for Transient Expression of Recombinant RNA
    6.
    发明申请
    Compositions and Methods for Transient Expression of Recombinant RNA 有权
    用于重组RNA瞬时表达的组合物和方法

    公开(公告)号：US20140249212A1

    公开(公告)日：2014-09-04

    申请号：US14343817

    申请日：2012-09-27

    申请人： Yale University

    发明人： Peter M. Rabinovich ,  Sherman M. Weissman

    IPC分类号： C12N15/85

    CPC分类号： C12N15/85 ,  C12N15/86 ,  C12N2760/18843 ,  C12N2760/18852

    摘要： Compositions for transient but prolonged exogenous mRNA expression through the use of the transcription system of negative strand RNA viruses, and methods of use thereof are disclosed. In some embodiments, the system contains only RNAs and does not include any DNA molecules. The compositions typically include an RNA template unit (rTeUn) that includes a virus regulatory sequences operably linked to a coding sequence of interest. The rTeUn is typically transfected to a host cell's cytoplasm in the presence of virus expression system proteins that mediate replication of the rTeUn and transcription of the transgene. The rTeUn RNA bonded to viral proteins exhibits high resistance to degradation, prolonged duration of expression, and is free of viral genes. The compositions can be used to reprogram cell. For example, the compositions and methods can be used to redirected lymphocytes to target cancer cells, or to dedifferentiate somatic cells into induce pluripotent stem cells.

    摘要翻译： 公开了通过使用负链RNA病毒的转录系统的瞬时但延长的外源mRNA表达的组合物及其使用方法。 在一些实施方案中，系统仅含有RNA并且不包括任何DNA分子。 组合物通常包括RNA模板单元（rTeUn），其包含与感兴趣的编码序列可操作地连接的病毒调节序列。 在介导rTeUn复制和转录转录的病毒表达系统蛋白的存在下，rTeUn通常被转染至宿主细胞的细胞质。 与病毒蛋白结合的rTeUn RNA表现出高度的降解抗性，延长的表达持续时间，并且没有病毒基因。 组合物可用于重新编程细胞。 例如，组合物和方法可用于将淋巴细胞重定向至靶癌细胞，或将体细胞去分化为诱导多能干细胞。