A high-throughput in vivo selection method for luciferase variants

This is the authors' abstract. We don't add key points for this paper.

Luciferase catalyzes its special substrate and light emits during the oxidation reaction, which is widely used as an optical sensor to monitor gene expression, protein-protein interaction and tumor response to therapy. However, limitations like the rapid decay, low stability and poor intensity of light emission make the optical signal hard to be captured in the sample detection. To obtain luciferase variants with improved features, traditional methods are usually based on CCD imaging technology or colony picking, which are low-throughput, time and labor-consuming. Herein, we describe a novel luciferase screening method called fast library shrink method (FLSM), which could efficiently help select the desired mutant from the huge and complicated pool without analyzing the variants one by one. Based on FLSM, we successfully obtained one Gluc variant named M43IL119Q, which shows a 3-fold increase in the intensity of light emission as well as a stable light emission kinetics. The selection method is ready-to-use and cost-effective, thus making it a promising platform for the fast evolution of luciferases.