Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Abdrabou, Abdalla M., Duong, Bill T. V., Chen, Kangfu, Atwal, Randy Singh, Labib, Mahmoud, Lin, Sichun, Angers, Stephane, Kelley, Shana O.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2022
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
_version_ 1784861505962901504
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
work_keys_str_mv AT abdrabouabdallam nuprismmicrofluidicgenomewidephenotypicscreeningplatformforcellularnuclei
AT duongbilltv nuprismmicrofluidicgenomewidephenotypicscreeningplatformforcellularnuclei
AT chenkangfu nuprismmicrofluidicgenomewidephenotypicscreeningplatformforcellularnuclei
AT atwalrandysingh nuprismmicrofluidicgenomewidephenotypicscreeningplatformforcellularnuclei
AT labibmahmoud nuprismmicrofluidicgenomewidephenotypicscreeningplatformforcellularnuclei
AT linsichun nuprismmicrofluidicgenomewidephenotypicscreeningplatformforcellularnuclei
AT angersstephane nuprismmicrofluidicgenomewidephenotypicscreeningplatformforcellularnuclei
AT kelleyshanao nuprismmicrofluidicgenomewidephenotypicscreeningplatformforcellularnuclei