Single-molecule fluorescence in situ hybridization (FISH) of circular RNA CDR1as
Authors
- C. Kocks
- A. Boltengagen
- M. Piwecka
- A. Rybak-Wolf
- N. Rajewsky
Journal
- Methods in Molecular Biology
Citation
- Methods Mol Biol 1724: 77-96
Abstract
Individual mRNA molecules can be imaged in fixed cells by hybridization with multiple, singly labeled oligonucleotide probes, followed by computational identification of fluorescent signals. This approach, called single-molecule RNA fluorescence in situ hybridization (smRNA FISH), allows subcellular localization and absolute quantification of RNA molecules in individual cells. Here, we describe a simple smRNA FISH protocol for two-color imaging of a circular RNA, CDR1as, simultaneously with an unrelated messenger RNA. The protocol can be adapted to circRNAs that coexist with overlapping, noncircular mRNA isoforms produced from the same genetic locus.