ENAapplication/xmlftp://ftp.sra.ebi.ac.uk/vol1/fastq/DRR525/DRR525842/DRR525842_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/DRR525/DRR525845/DRR525845_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/DRR525/DRR525846/DRR525846_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/DRR525/DRR525844/DRR525844_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/DRR525/DRR525845/DRR525845_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/DRR525/DRR525847/DRR525847_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/DRR525/DRR525841/DRR525841_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/DRR525/DRR525844/DRR525844_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/DRR525/DRR525847/DRR525847_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/DRR525/DRR525843/DRR525843.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/DRR525/DRR525842/DRR525842_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/DRR525/DRR525846/DRR525846_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/DRR525/DRR525841/DRR525841_1.fastq.gzprimaryOK200THP1 cellsGenomicsThe University of Tokyohttps://www.ebi.ac.uk/ena/browser/view/PRJDB17362Homo sapiensRecent advancements in image-based pooled CRISPR screening have facilitated the mapping of diverse genotype-phenotype associations within mammalian cells. However, the rapid enrichment of cells based on morphological information continues to pose a challenge, thereby constraining the capacity for large-scale gene perturbation screening across diverse high-content cellular phenotypes. In this study, we demonstrate the applicability of multimodal ghost cytometry-based cell sorting, including both fluorescent and label-free high-content phenotypes, for rapid pooled CRISPR screening within vast cell populations. Using the high-content cell sorter operating in fluorescence mode, we successfully executed kinase-specific CRISPR screening targeting genes influencing the nuclear translocation of RelA. Furthermore, using the multi-parametric, label-free mode, we performed large-scale screening to identify genes involved in macrophage polarization. Notably, the label-free platform can enrich target phenotypes without requiring invasive staining, preserving untouched cells for downstream assays, and expanding the potential for screening cellular phenotypes even when suitable markers are absent.xref:EuropePMC:PMC10985231ENAfalseHomo sapiens strain:THP1 cellsPooled CRISPR Screening of High-content Cellular Phenotypes by Ghost Cytometry2025-09-242024-01-26PRJDB173629606