Solid phase chemistry to covalently and reversibly capture thiolated RNA

• Published in: Nucleic acids research Vol. 46; no. 14; pp. 6996 - 7005
• Main Authors: Duffy, Erin E, Canzio, Daniele, Maniatis, Tom, Simon, Matthew D
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 21.08.2018
• Subjects: Animals; Cell Line, Tumor; Chemical Biology and Nucleic Acid Chemistry; Gene Expression - genetics; HEK293 Cells; Humans; Mesylates - chemistry; Mice; MicroRNAs - genetics; Neurons - cytology; RNA - chemistry; RNA - genetics; RNA - metabolism; RNA Polymerase II - metabolism; Staining and Labeling - methods; Thiouridine - chemistry

Abstract Here, we describe an approach to enrich newly transcribed RNAs from primary mouse neurons using 4-thiouridine (s4U) metabolic labeling and solid phase chemistry. This one-step enrichment procedure captures s4U-RNA by using highly efficient methane thiosulfonate (MTS) chemistry in an immobil...