Genome-scale requirements for dynein-based transport revealed by a high-content arrayed CRISPR screen

Abstract

The microtubule motor dynein plays a key role in cellular organization. However, little is known about how dynein’s biosynthesis, assembly, and functional diversity are orchestrated. To address this issue, we have conducted an arrayed CRISPR loss-of-function screen in human cells using the distribution of dynein-tethered peroxisomes and early endosomes as readouts. From a genome-wide gRNA library, 195 validated hits were recovered and parsed into those impacting multiple dynein cargoes and those whose effects are restricted to a subset of cargoes. Clustering of high-dimensional phenotypic fingerprints revealed co-functional proteins involved in many cellular processes, including several candidate novel regulators of core dynein functions. Further analysis of one of these factors, the RNA-binding protein SUGP1, indicates that it promotes cargo trafficking by sustaining functional expression of the dynein activator LIS1. Our data represent a rich source of new hypotheses for investigating microtubule-based transport, as well as several other aspects of cellular organization captured by our high-content imaging.

Spatiotemporal proteomic profiling of cellular responses to NLRP3 agonists

“…However, our data support a model where TGN38/46 dispersion into endosomes is a consequence of cellular stress rather than a feature of NLRP3 activation, in agreement with recent studies [58][59]. Further supporting this conclusion, a phenotypic CRISPR screen demonstrated that many cellular perturbations of transport likely unrelated to NLRP3 activation induce TGN38/46 peripheral redistribution [171].…”