Global Run-On sequencing to measure nascent transcription in C. elegans

STAR Protoc. 2021 Dec 4;2(4):100991. doi: 10.1016/j.xpro.2021.100991. eCollection 2021 Dec 17.

Abstract

Global Run-On sequencing (GRO-seq) is one of the most sensitive techniques to detect nascent transcription from RNA polymerase (Pol) at a genome-wide level. The protocol incorporates labeled ribonucleotides into nascent RNAs from Pol I, II, and III. We have adapted the GRO-seq protocol to the nematode Caenorhabditis elegans to measure transcription from embryos and adult worms. Here, we provide a detailed overview of the protocol highlighting the critical steps for generating successful libraries. For complete details on the use and execution of this protocol, please refer to Quarato et al. (2021).

Keywords: Cell separation/fractionation; Gene Expression; Genomics; Model Organisms; Molecular Biology; RNAseq; Sequencing.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Caenorhabditis elegans* / genetics
  • Caenorhabditis elegans* / metabolism
  • Cell Nucleus / chemistry
  • Gene Library
  • Immunoprecipitation / methods
  • RNA, Helminth* / analysis
  • RNA, Helminth* / genetics
  • Sequence Analysis, RNA / methods*
  • Transcription, Genetic / genetics*

Substances

  • RNA, Helminth