nuPRISM: Microfluidic Genome-Wide Phenotypic Screening Platform for Cellular Nuclei

  • ACS Cent Sci. 2022 Dec 28;8(12):1618-1626. doi: 10.1021/acscentsci.2c00836.
Abdalla M Abdrabou  1 ,  Bill T V Duong  2 ,  Kangfu Chen  2 ,  Randy Singh Atwal  1 ,  Mahmoud Labib  3 ,  Sichun Lin  4 ,  Stephane Angers  2  5  4 ,  Shana O Kelley  1  3  6  2
Affiliations
  • 1. Department of Biochemistry and Molecular Genetics, Feinberg School of Medicine, Northwestern University, Chicago, Illinois 60611, United States.
  • 2. Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, Ontario M5S 3M2, Canada.
  • 3. Department of Chemistry, Northwestern University, Evanston, Illinois 60611, United States.
  • 4. Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario M5S 3E1, Canada.
  • 5. Department of Biochemistry, Faculty of Medicine, University of Toronto, Toronto, Ontario M5S 1A8, Canada.
  • 6. Department of Biomedical Engineering, Northwestern University, Evanston, Illinois 60611, United States.
PMID: 36589880 DOI: 10.1021/acscentsci.2c00836
Abstract

Genome-wide loss-of-function screens are critical tools to identify novel genetic regulators of intracellular proteins. However, studying the changes in the organelle-specific expression profile of intracellular proteins can be challenging due to protein localization differences across the whole cell, hindering context-dependent protein expression and activity analyses. Here, we describe nuPRISM, a microfluidics chip specifically designed for large-scale isolated nuclei sorting. The new device enables rapid genome-wide loss-of-function phenotypic CRISPR-Cas9 screens directed at intranuclear targets. We deployed this technology to identify novel genetic regulators of β-catenin nuclear accumulation, a phenotypic hallmark of APC-mutated colorectal Cancer. nuPRISM expands our ability to capture aberrant nuclear morphological and functional traits associated with distinctive signal transduction and subcellular localization-driven functional processes with substantial resolution and high throughput.

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