In situ genotyping of a pooled strain library after characterizing complex phenotypes

In this work, we present a proof-of-principle experiment that extends advanced live cell microscopy to the scale of pool-generated strain libraries. We achieve this by identifying the genotypes for individual cells in situ after a detailed characterization of the phenotype. The principle is demonstrated by single-molecule fluorescence time-lapse imaging of Escherichia coli strains harboring barcoded plasmids that express a sgRNA which suppresses different genes in the E. coli genome through dCas9 interference. In general, the method solves the problem of characterizing complex dynamic phenotypes for diverse genetic libraries of cell strains. For example, it allows screens of how changes in regulatory or coding sequences impact the temporal expression, location, or function of a gene product, or how the altered expression of a set of genes impacts the intracellular dynamics of a labeled reporter.

Source: Molecular Systems Biology - October 17, 2017 Category: Molecular Biology Authors: Lawson, M. J., Camsund, D., Larsson, J., Baltekin, O., Fange, D., Elf, J. Tags: Methods & Resources, Quantitative Biology & Dynamical Systems Source Type: research