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nuPRISM: Microfluidic Genome-Wide Phenotypic Screening Platform for Cellular Nuclei
[Image: see text] 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 ac...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2022
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9801500/ https://www.ncbi.nlm.nih.gov/pubmed/36589880 http://dx.doi.org/10.1021/acscentsci.2c00836 |
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author | Abdrabou, Abdalla M. Duong, Bill T. V. Chen, Kangfu Atwal, Randy Singh Labib, Mahmoud Lin, Sichun Angers, Stephane Kelley, Shana O. |
author_facet | Abdrabou, Abdalla M. Duong, Bill T. V. Chen, Kangfu Atwal, Randy Singh Labib, Mahmoud Lin, Sichun Angers, Stephane Kelley, Shana O. |
author_sort | Abdrabou, Abdalla M. |
collection | PubMed |
description | [Image: see text] 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. |
format | Online Article Text |
id | pubmed-9801500 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-98015002022-12-31 nuPRISM: Microfluidic Genome-Wide Phenotypic Screening Platform for Cellular Nuclei Abdrabou, Abdalla M. Duong, Bill T. V. Chen, Kangfu Atwal, Randy Singh Labib, Mahmoud Lin, Sichun Angers, Stephane Kelley, Shana O. ACS Cent Sci [Image: see text] 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. American Chemical Society 2022-11-28 2022-12-28 /pmc/articles/PMC9801500/ /pubmed/36589880 http://dx.doi.org/10.1021/acscentsci.2c00836 Text en © 2022 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Abdrabou, Abdalla M. Duong, Bill T. V. Chen, Kangfu Atwal, Randy Singh Labib, Mahmoud Lin, Sichun Angers, Stephane Kelley, Shana O. nuPRISM: Microfluidic Genome-Wide Phenotypic Screening Platform for Cellular Nuclei |
title | nuPRISM: Microfluidic
Genome-Wide Phenotypic Screening
Platform for Cellular Nuclei |
title_full | nuPRISM: Microfluidic
Genome-Wide Phenotypic Screening
Platform for Cellular Nuclei |
title_fullStr | nuPRISM: Microfluidic
Genome-Wide Phenotypic Screening
Platform for Cellular Nuclei |
title_full_unstemmed | nuPRISM: Microfluidic
Genome-Wide Phenotypic Screening
Platform for Cellular Nuclei |
title_short | nuPRISM: Microfluidic
Genome-Wide Phenotypic Screening
Platform for Cellular Nuclei |
title_sort | nuprism: microfluidic
genome-wide phenotypic screening
platform for cellular nuclei |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9801500/ https://www.ncbi.nlm.nih.gov/pubmed/36589880 http://dx.doi.org/10.1021/acscentsci.2c00836 |
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