Extended Data Fig. 1: Overview of magnetic ranking cytometry (MagRC)-based cell sorting and endogenous KRAS protein level-dependent gRNA enrichment analysis of pooled genome-wide CRISPR-edited cells.

a, Schematic design of the microfluidic chip utilized for high-throughput cell sorting of TKOv3 CRISPR KO gRNA library transduced cells. The fluidic channel incorporates X-shaped microfabricated structures of different heights forming capture pockets to trap the magnetic nanoparticle labeled cells. Cells carrying high levels of immunomagnetic nanoparticles acquire a stronger magnetic force that can overcome the fluidic drag forces. Cells captured in zone 1 have high KRAS protein expression, and those in zone 2 have medium KRAS expression. The outlet connects to a syringe for the collection of cells with low KRAS expression. b, Representative flow cytometry profile of KRAS protein expression after sorting and cell sorting profiles of the screened cells (200 million cells per replicate). c, d, Scatter plots of individual gene enrichment scores and fold change over the unsorted population for (c) WT KRAS-expressing HT29 and (d) homozygous G12V KRAS-expressing SW480 cells.