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nuPRISM: Microfluidic Genome-Wide Phenotypic Screening Platform for Cellular Nuclei
Abdalla M Abdrabou1, Bill T V Duong2, Kangfu Chen2
1Department of Biochemistry and Molecular Genetics, Feinberg School of Medicine, Northwestern University, Chicago, Illinois 60611, United States.
ACS Central Science
|January 2, 2023
Summary
We developed nuPRISM, a microfluidics chip for high-throughput CRISPR screens targeting intranuclear proteins. This tool aids in discovering genetic regulators of nuclear processes, like β-catenin accumulation in colorectal cancer.
Area of Science:
- Cell Biology
- Genomics
- Biotechnology
Background:
- Genome-wide screens are vital for identifying genetic regulators of intracellular proteins.
- Analyzing organelle-specific protein expression is challenging due to localization complexities.
- Context-dependent protein activity analysis requires precise localization studies.
Purpose of the Study:
- To introduce nuPRISM, a microfluidics chip for large-scale isolated nuclei sorting.
- To enable genome-wide loss-of-function CRISPR-Cas9 screens targeting intranuclear proteins.
- To identify novel genetic regulators of β-catenin nuclear accumulation in colorectal cancer.
Main Methods:
- Development of nuPRISM, a microfluidics chip for nuclei sorting.
- Application of CRISPR-Cas9 for genome-wide loss-of-function screens.
- High-throughput screening of intranuclear targets.
Main Results:
- nuPRISM facilitates rapid, genome-wide screens focused on intranuclear targets.
- The technology successfully identified novel regulators of β-catenin nuclear accumulation.
- Enabled capture of nuclear morphological and functional traits with high resolution.
Conclusions:
- nuPRISM enhances the study of intranuclear protein regulation.
- The chip provides a powerful tool for dissecting signal transduction and localization-driven processes.
- Facilitates high-throughput analysis of nuclear phenotypes in diseases like colorectal cancer.
