Year: 2018
Type: article
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 immobilized format. Like solution-based methods, this solid-phase enrichment can distinguish mature RNAs (mRNA) with differential stability, and can be used to reveal transient RNAs such as enhancer RNAs (eRNAs) and primary microRNAs (pri-miRNAs) from short metabolic labeling. Most importantly, the efficiency of this solid-phase chemistry made possible the first large scale measurements of RNA polymerase II (RNAPII) elongation rates in mouse cortical neurons. Thus, our approach provides the means to study regulation of RNA metabolism in specific tissue contexts as a means to better understand gene expression in vivo.
Source: Nucleic Acids Research
DOI: 10.1093/nar/gky556
Institutions: Yale University, Columbia University Irving Medical Center
Language: English
Topic: RNA Research and Splicing
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,992
Awards: Predoctoral Program in Cellular and Molecular Biology, Graduate Research Fellowship Program, Integrating RNAs into signaling pathways by engineering covalent RNA modification, The role of the clustered protocadherins in neural circuit formation and implications for neurodevelopmental disorders
OpenAlex ID: W2833484261