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

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.