Labeling of Nascent RNA in C. elegans Intestine
This article has been Reviewed by the following groups
Discuss this preprint
Start a discussion What are Sciety discussions?Listed in
- Evaluated articles (Arcadia Science)
Abstract
Transcriptional regulation in C. elegans has been difficult to study at the level of nascent RNA because nucleotide analogs do not readily penetrate the cuticle. Here, we establish an ex vivo EU-click labeling protocol that enables sensitive microscopy detection of newly transcribed RNA in dissected intestines. Using worms expressing fluorescent nucleolar markers, we show that EU incorporation faithfully reports on rRNA nascent transcription and is abolished by RNA polymerase I inhibition. Spatial analysis further reveals that the majority of nascent rRNA transcripts localize to the fibrillar zone (FZ) of intestinal nucleoli, consistent with the conserved role of this compartment in rRNA synthesis. In addition to imaging applications, this workflow can be adapted for gene expression assays, providing a versatile approach for quantitative analysis of nascent transcription in C. elegans . By enabling direct visualization of nucleolar transcription in intact intestine tissue, this method opens new opportunities to investigate how nucleolar activity is regulated across development, aging, and disease contexts.
Article activity feed
-
By combining EU labeling with compartment-specific markers, we find that the EU signal localizes to a reticulated network within the fibrillar zone, contrasting with the discrete transcriptional foci often reported in cultured mammalian cells
I'm curious if you have compared this method to approaches like live imaging of MS2 RNA hairpin loops?
-
Together, these results establish a robust and versatile method for visualizing and quantifying nascent transcription in intact C. elegans tissues.
This is a great, new methodology to study nascent transcription in C. elegans! I'm looking forward to seeing how this approach is extended to incorporate quantitative approaches like EU-RNA-seq or qPCR.
-
