title: Single-cell detection of trans-splicing ribozyme in vivo activity.
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.
authors.affiliation: Department of Radiology & Bio-X Program, Molecular Imaging Program at Stanford, Stanford University, 300 Pasteur Drive, Stanford, California 94305-5344, USA.
authors.forename: Sumitaka
authors.initials: S
authors.lastname: Hasegawa
authors.forename: Jung W
authors.initials: JW
authors.lastname: Choi
authors.forename: Jianghong
authors.initials: J
authors.lastname: Rao
mesh_terms: D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D000882:Haplorhini; D008856:Microscopy, Fluorescence; D016337:RNA, Catalytic / Q000302:isolation & purification* / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020040:Trans-Splicing
chemical_list: D016337:RNA, Catalytic; D012333:RNA, Messenger