Problem
Ribosome profiling enables the study of translation and translational control across the transcriptome. Despite intensive efforts since initial publication of the methodology in 2009, this experimental technique remains restricted in terms of applicability, typically requiring ~1 million cells as input. No current commercially available technology is capable of measuring translation in single cells.
Solution
Dr. Can Cenik at The University of Texas at Austin has adapted a microfluidics chip method that leverages the advantages of IsoTachoPhoresis (ITP) to enable ribosomal profiling in extremely low input samples volumes including single cells. ITP offers faster processing times, no requirement of liquid transfers, and high yield with low RNA inputs. Dr. Cenik has optimized ITP methods to introduce highly stringent size selection to enable ribosome profiling applications. This technique significantly reduces input sample requirements for ribosomal profiling by many orders of magnitude and introduces highly stringent size selection in order to deliver ribosome occupancy measurements from ultra-low input samples, including single cells.
Benefits
- Enables single cell and ultra-low input ribosomal profiling
- Validated in single oocytes, human cancer cells, and mouse embryonic cells
- Simple method couples IsoTachoPhoresis with polyacrylamide gel based size selection
Publication
Ozadam et al. 2023 Nature: https://www.nature.com/articles/s41586-023-06228-9
