2. Academic Validation
  3. Isolation of Newly Transcribed RNA Using the Metabolic Label 4-Thiouridine

Isolation of Newly Transcribed RNA Using the Metabolic Label 4-Thiouridine

  • Methods Mol Biol. 2017:1648:169-176. doi: 10.1007/978-1-4939-7204-3_13.
Angela Garibaldi 1,  Francisco Carranza 1,  Klemens J Hertel 2
Affiliations

Affiliations

  • 1 Department of Microbiology and Molecular Genetics, University of California, Irvine, B233 Med Sci I, Irvine, CA, 92697-4025, USA.
  • 2 Department of Microbiology and Molecular Genetics, University of California, Irvine, B233 Med Sci I, Irvine, CA, 92697-4025, USA. khertel@uci.edu.
PMID: 28766297 DOI: 10.1007/978-1-4939-7204-3_13
Abstract

Isolation of newly transcribed RNA is an invaluable approach that can be used to study the dynamic life of RNA in cellulo. Traditional methods of whole-cell RNA extraction limit subsequent gene expression analyses to the steady-state levels of RNA abundance, which often masks changes in RNA synthesis and processing. This chapter describes a methodology with low cytotoxicity that permits the labeling and isolation of nascent pre-mRNA in Cell Culture. The resulting isolate is suitable for use in a series of downstream applications aimed at studying changes in RNA synthesis, processing, or stability.

Keywords

4-Thiouridine; 4sU; 4sU-seq; Decay; Mammalian cells; Metabolic labeling; Nascent RNA; Nascent pre-mRNA; Transcription; mRNA processing.

Figures
Products