Single-cell detection of trans-splicing ribozyme in vivo activity.
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.
Hasegawa
Sumitaka
S
Department of Radiology & Bio-X Program, Molecular Imaging Program at Stanford, Stanford University, 300 Pasteur Drive, Stanford, California 94305-5344, USA.
Choi
Jung W
JW
Rao
Jianghong
J
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
D016337:RNA, Catalytic; D012333:RNA, Messenger