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Pooled CRISPR screening of high-content cellular phenotypes by ghost cytometry
Pooled CRISPR screening of high-content cellular phenotypes by ghost cytometry
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Length:
20 minutes
Released:
Jan 27, 2023
Format:
Podcast episode
Description
Link to bioRxiv paper:
http://biorxiv.org/cgi/content/short/2023.01.26.525784v1?rss=1
Authors: Tsubouchi, A., An, Y., Kawamura, Y., Yanagihashi, Y., Murata, Y., Teranishi, K., Ishiguro, S., Aburatani, H., Yachie, N., Ota, S.
Abstract:
Fast enrichment of cells based on morphological information remains a challenge, limiting genome-wide perturbation screening for diverse high-content phenotypes of cells. Here we show that multi-modal ghost cytometry-based cell sorting is applicable to fast pooled CRISPR screening for both fluorescence and label-free high-content phenotypes of millions of cells. By employing the high-content cell sorter in the fluorescence mode, we enabled the genome-wide CRISPR screening of genes that affect NF-{kappa}B nuclear translocation. Furthermore, by employing the multi-parametric, label-free mode, we performed the large-scale screening to identify a gene involved in macrophage polarization. Especially the label-free platform can enrich target phenotypes without invasive staining, preserving untouched cells for downstream assays and unlocking the potential to screen for the cellular phenotypes even when suitable markers are lacking.
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Podcast created by Paper Player, LLC
http://biorxiv.org/cgi/content/short/2023.01.26.525784v1?rss=1
Authors: Tsubouchi, A., An, Y., Kawamura, Y., Yanagihashi, Y., Murata, Y., Teranishi, K., Ishiguro, S., Aburatani, H., Yachie, N., Ota, S.
Abstract:
Fast enrichment of cells based on morphological information remains a challenge, limiting genome-wide perturbation screening for diverse high-content phenotypes of cells. Here we show that multi-modal ghost cytometry-based cell sorting is applicable to fast pooled CRISPR screening for both fluorescence and label-free high-content phenotypes of millions of cells. By employing the high-content cell sorter in the fluorescence mode, we enabled the genome-wide CRISPR screening of genes that affect NF-{kappa}B nuclear translocation. Furthermore, by employing the multi-parametric, label-free mode, we performed the large-scale screening to identify a gene involved in macrophage polarization. Especially the label-free platform can enrich target phenotypes without invasive staining, preserving untouched cells for downstream assays and unlocking the potential to screen for the cellular phenotypes even when suitable markers are lacking.
Copy rights belong to original authors. Visit the link for more info
Podcast created by Paper Player, LLC
Released:
Jan 27, 2023
Format:
Podcast episode
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