Single-cell detection of trans-splicing ribozyme in vivo activity |
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Authors: | Hasegawa Sumitaka Choi Jung W Rao Jianghong |
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Affiliation: | Department of Radiology & Bio-X Program, Molecular Imaging Program at Stanford, Stanford University, 300 Pasteur Drive, Stanford, California 94305-5344, USA. |
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Abstract: | The Tetrahymena trans-splicing ribozyme can edit RNA in a sequence-specific manner, but its efficiency needs to be improved for any functional rescues. This communication describes a simple method that uses a bacterial enzyme beta-lactamase to report trans-splicing activity of Tetrahymena ribozyme in single living mammalian cells by fluorescence microscopy and flow cytometry. This enzyme-based single-cell detection method is highly sensitive and compatible with living cell flow cytometry, and should allow a cell-based systematic screening of a vast library of ribozymes for better trans-spliced ribozyme variants. |
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