摘要:
Methods and compositions for producing hRNA, e.g., shRNA, expression modules for specific target nucleic acids are provided. In the subject methods, an initial nucleic acid, e.g., dsDNA, synthetic DNA, etc., corresponding to the target nucleic acid of interest is converted to an intermediate nucleic acid. The resultant intermediate nucleic acid, following an optional size modification step, is then converted to a linear dsDNA that includes at least one copy of the hRNA expression module of interest, or a precursor (i.e., pro-shRNA expression module) thereof, where in certain embodiments conversion may include amplification. Also provided are reagents, systems and kits for use in practicing the subject methods. The subject methods and compositions find use in a variety of different applications.
摘要:
Sequential reporter enzyme luminescence (SRL) methods are provided. In the subject methods, the activity of a reporter enzyme is evaluated using a secondary reporter system that employs a product of a reporter enzyme mediated reaction as a luminescent substrate, e.g., luciferase substrate. Also provided are kits and other compositions that find use in practicing the subject methods.
摘要:
Methods and compositions for detecting molecular interactions are provided. Aspects of the invention include the use of a reduced affinity enzyme complementation reporter system. Also provided are systems and kits for use in practicing embodiments of the methods.
摘要:
Methods and compositions are provided for effecting physiological processes using a conjugate of a binding member binding to a surface membrane marker and a ligand for a cell surface receptor, where the ligand is attenuated as compared to the natural ligand. The composition finds particular application in discriminating between cells that share the receptor and can be distinguished by the surface membrane marker.
摘要:
Methods and compositions for detecting the sub-cellular localization of a molecule are provided. Aspects of the invention include detecting translocation of a cell-surface receptor to a sub-cellular compartment, e.g., the endosome, using a reduced affinity enzyme complementation reporter system. Also provided are systems and kits for use in practicing embodiments of the methods.
摘要:
Methods and compositions for detecting molecular interactions are provided. Aspects of the invention include the use of a reduced affinity enzyme complementation reporter system. Also provided are systems and kits for use in practicing embodiments of the methods.
摘要:
Methods and compositions for detecting the sub-cellular localization of a molecule are provided. Aspects of the invention include detecting translocation of a cell-surface receptor to a sub-cellular compartment, e.g., the endosome, using a reduced affinity enzyme complementation reporter system. Also provided are systems and kits for use in practicing embodiments of the methods.
摘要:
Sequential reporter enzyme luminescence (SRL) methods are provided. In the subject methods, the activity of a reporter enzyme is evaluated using a secondary reporter system that employs a product of a reporter enzyme mediated reaction as a luminescent substrate, e.g., luciferase substrate. Also provided are kits and other compositions that find use in practicing the subject methods.