Riboseq

Riboseq, also known as ribosome profiling, is a high-throughput sequencing technique used to analyze the positions of ribosomes on messenger RNA (mRNA) molecules within living cells. This method provides insights into translation dynamics by capturing snapshots of actively translating ribosomes, thereby revealing where protein synthesis occurs and how frequently specific mRNAs are being translated.

The process of Riboseq involves treating cells with nucleases that digest unprotected regions of mRNA, leaving behind fragments protected by ribosomes, known as ribosome-protected footprints. These fragments are then isolated, converted into cDNA libraries, and subjected to next-generation sequencing. The resulting data allows researchers to map ribosome positions at nucleotide resolution across the transcriptome.

Riboseq enables the quantification of translation efficiency, identification of initiation sites, and analysis of translational regulation under different physiological or environmental conditions. It has advanced understanding in fields such as gene expression regulation, cellular response mechanisms, and disease pathology, including cancer and viral infections.

Since its development in the early 2010s, Riboseq has become a critical tool in molecular biology and systems biology research. It provides a detailed view of the translational landscape, complementing transcriptomics data, and helps elucidate the complex relationship between mRNA abundance and protein production.

Overall, Riboseq is a powerful and precise technique for studying protein synthesis in vivo, contributing to a deeper understanding of gene regulation at the translational level. Its applications continue to grow with ongoing improvements in sequencing technologies and analytical methods.