Visualization of induced RNA in single bacterial cells

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 714; p. 189
• Main Authors: Borogovac, Azra, Broude, Natalia E
• Format: Journal Article
• Language: English
• Published: United States 2011
• Subjects: Aptamers, Nucleotide - metabolism; Cells, Immobilized - cytology; Cells, Immobilized - metabolism; Escherichia coli - cytology; Escherichia coli - genetics; Escherichia coli - metabolism; Flow Cytometry; Genes, Reporter - genetics; Glass - chemistry; Kinetics; Microscopy, Fluorescence; Molecular Imaging - methods; Plasmids - genetics; Protein Biosynthesis; RNA, Bacterial - analysis; RNA, Bacterial - biosynthesis; RNA, Bacterial - chemistry; RNA, Bacterial - genetics; Single-Cell Analysis - methods; Surface Properties; Transcription, Genetic
• ISSN: 1940-6029
• DOI: 10.1007/978-1-61779-005-8_12

Visualization of RNA in live cells is a challenging task due to the transient character of most RNA molecules and the lack of adequate methods to label RNA noninvasively. Here, we describe a system for regulated RNA synthesis and visualization of RNA in live Escherichia coli cells based on protein complementation. This method allows for labeling RNA with a relatively small protein complex that becomes fluorescent only when bound to an RNA. This method greatly reduces the high fluorescence background characteristic of methods employing intact fluorescent proteins. A short reporter RNA was shown to localize at the cell periphery in nonrandom patterns.