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    • 2. 发明申请
    • FLUORESCENT BIOSENSOR FOR 2', 3'-cGAMP
    • 荧光生物传感器2',3'-cGAMP
    • WO2017218358A1
    • 2017-12-21
    • PCT/US2017/036859
    • 2017-06-09
    • THE REGENTS OF THE UNIVERSITY OF CALIFORNIA
    • HAMMOND, Ming C.SU, YichiBOSE, Debojit
    • A61K31/7125A61K31/713C12N15/113C12N15/115C12Q1/68G01N33/53
    • C12N15/113A61K31/7105A61K31/713C12N15/115C12N2310/16C12N2320/10C12Q1/6825
    • A single stranded nucleic acid biosensor for 2', 3'-cGAMP is provided. The single stranded nucleic acid may include a 2', 3'-cGAMP-binding riboswitch domain comprising a transducer stem and a dye-binding aptamer domain that is operably connected to the transducer stem. A 2', 3'-cGAMP-binding riboswitch domain can be a GEMM-II riboswitch domain. The dye-binding aptamer domain can be a Spinach2 aptamer. The 2', 3'-cGAMP biosensor may further include a signaling chromophore specifically bound to the Spinach2 aptamer domain, where the sensor is configured to fluorescently activate the signaling chromophore upon specific binding of 2', 3'-cGAMP to the 2', 3'-cGAMP-binding riboswitch domain. Also provided are methods in which the subject 2', 3'-cGAMP biosensors find use including methods for determining the level of cGAS activity in a sample or a cell. Nucleic acid constructs and host cells including the same are also provided.
    • 提供了用于2',3'-cGAMP的单链核酸生物传感器。 单链核酸可包括2',3'-cGAMP-结合核糖开关结构域,其包含可操作地连接至换能器茎的换能器茎和染料结合适体结构域。 2',3'-cGAMP结合核糖开关结构域可以是GEMM-II核糖开关结构域。 染料结合适体结构域可以是Spinach2适体。 2',3'-cGAMP生物传感器可以进一步包括与Spinach2适体结构域特异性结合的信号发色团,其中传感器被配置为在2',3'-cGAMP与2',3'-cGAMP特异性结合后荧光激活信号发色团, 3'-cGAMP-结合核糖开关结构域。 还提供了其中受试者2',3'-cGAMP生物传感器的使用方法,包括用于确定样品或细胞中cGAS活性水平的方法。 还提供了核酸构建体和包括它们的宿主细胞。