The use of Benzonase to produce ribosome footprints simplifies translational levels quantification by Ribo-seq.

ABSTRACT: Gene expression quantification through genomics methods is crucial for understanding diverse biological contexts. Among these methods, ribosome profiling (Ribo-seq) stands out as a valuable tool for uncovering post-transcriptional gene expression regulation by providing a comprehensive view of the translatome. While current protocols are time-intensive with limited variations, we introduced the use of the Benzonase enzyme to generate ribosome footprints from a polysome-enriched fraction that exhibit expected characteristics in size, transcriptome mapping, and periodicity. Comparing translatome from Benzonase- and RNAse I-derived footprints reveals minimal differences underscoring Benzonase's potential to streamline the protocol, reducing time, cost and bias. We further demonstrate Ribo-seq

SUBMITTER: Eastman G 

PROVIDER: S-EPMC11952333 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature