Systems and methods for measuring translation activity in viable cells

摘要

Systems for measuring protein translation and methods for measuring overall translation activity in viable cells or subcellular compartments is disclosed. The methods identify general ribosomal activity, if desired at sub-cellular resolution, thereby providing a signal indicating the rate of any of the steps of protein synthesis selected from initiation, elongation, termination or recycling. The translation system can be used to identify translation modulators in high-throughput-screening (HTS).

主权项

1. A method for measuring translation activity, which comprises:
labeling at least one tRNA with a first label and at least one tRNA with a second label, wherein when the first label and the second label are in proximity a detectable electromagnetic radiation is produced; introducing into a biological sample the at least one tRNA labeled with the first label and the at least one tRNA labeled with the second label, wherein said labels produce electromagnetic radiation in response to overall protein translation activity in said biological sample, wherein said biological sample is an intact cell; detecting said electromagnetic radiation from the sample; analyzing said electromagnetic radiation to produce a readout of a parameter of translation activity, wherein the parameter is selected from the group consisting of rate of translational activity, ratio of labeled versus unlabeled tRNA, location of translation activity, and average translation speed; and comparing said readout to a reference standard thereby determining an estimate of overall translation activity in real time without identifying a protein synthesized.