Open AccessIn situ genotyping of a pooled strain library after characterizing complex phenotypes
Author(s) -
Lawson Michael J,
Camsund Daniel,
Larsson Jimmy,
Baltekin Özden,
Fange David,
Elf Johan
Publication year - 2017
Publication title -
molecular systems biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 8.523
H-Index - 148
ISSN - 1744-4292
DOI - 10.15252/msb.20177951
Subject(s) - biology , library science , cell and molecular biology , molecular cell biology , genetics , gene , computer science , microbiology and biotechnology , plant development
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 sg RNA which suppresses different genes in the E. coli genome through dC as9 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.