Active Ribosome Profiling

^{Unveiling the Actively Translated Population}

How it compares to standard Ribo-Seq

A population of messenger RNA molecules exists in a "stalled" state. In this scenario, ribosomes are loaded onto the mRNA and ready for translation, but actual protein production does not occur.
Standard Ribo-Seq captures all ribosomes on mRNA, including those paused and not actively translating proteins. This underestimates true translation efficiency and weakens the connection to protein levels measured by proteomics methodologies. 
Active Ribo-Seq only targets actively translating ribosomes. This allows the discrimination of the active and the stalled ribosome populations and offers a more accurate picture of translation efficiency.

Conventional Ribo-Seq techniques rely on separating ribosomes based on their association with mRNA (polysomes) using fractionation on a density gradient. Since actively translating and stalled ribosomes have the same molecular weight, this method can't differentiate between them. 
RiboLace^TM is Immagina's proprietary technique that selects ribosomes based on their conformational state. By relying on a probe that is specific for ribosomes in the open state, it is possible to specifically isolate and analyze ribosomes that are actively translating mRNA.