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In situ genotyping of a pooled strain library after characterizing complex phenotypes
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Molecular Systems Biology. Uppsala University, Science for Life Laboratory, SciLifeLab.
Uppsala University, Science for Life Laboratory, SciLifeLab. Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Molecular Systems Biology.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Molecular Systems Biology. Uppsala University, Science for Life Laboratory, SciLifeLab.
Uppsala University, Science for Life Laboratory, SciLifeLab. Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Molecular Systems Biology.
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2017 (English)In: Molecular Systems Biology, ISSN 1744-4292, E-ISSN 1744-4292, Vol. 13, no 10, article id 947Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
2017. Vol. 13, no 10, article id 947
Keywords [en]
DuMPLING, live cell, microfluidic, single cell, strain libraries
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-342924DOI: 10.15252/msb.20177951ISI: 000416160000004PubMedID: 29042431OAI: oai:DiVA.org:uu-342924DiVA, id: diva2:1185625
Funder
Knut and Alice Wallenberg FoundationSwedish Research CouncilEU, European Research CouncilAvailable from: 2018-02-26 Created: 2018-02-26 Last updated: 2018-02-26Bibliographically approved

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