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

Lawson MJ, Camsund D, Larsson J, Baltekin Ö, Fange D, Elf J

Mol Syst Biol 13 (10) 947 [2017-10-17; online 2017-10-17]

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.

Affiliated researcher

PubMed 29042431

DOI 10.15252/msb.20177951

Crossref 10.15252/msb.20177951

pmc: PMC5658705

Publications 9.5.0