High-content imaging-based pooled CRISPR screens in mammalian cells
CRISPR (clustered regularly interspaced short palindromic repeats)-based gene inactivation provides a powerful means for linking genes to particular cellular phenotypes. CRISPR-based screening typically uses large genomic pools of single guide RNAs (sgRNAs). However, this approach is limited to phen...
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| Published in | The Journal of cell biology Vol. 220; no. 2 |
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| Main Authors | , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
United States
Rockefeller University Press
01.02.2021
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| Subjects | |
| Online Access | Get full text |
| ISSN | 0021-9525 1540-8140 1540-8140 |
| DOI | 10.1083/jcb.202008158 |
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| Abstract | CRISPR (clustered regularly interspaced short palindromic repeats)-based gene inactivation provides a powerful means for linking genes to particular cellular phenotypes. CRISPR-based screening typically uses large genomic pools of single guide RNAs (sgRNAs). However, this approach is limited to phenotypes that can be enriched by chemical selection or FACS sorting. Here, we developed a microscopy-based approach, which we name optical enrichment, to select cells displaying a particular CRISPR-induced phenotype by automated imaging-based computation, mark them by photoactivation of an expressed photoactivatable fluorescent protein, and then isolate the fluorescent cells using fluorescence-activated cell sorting (FACS). A plugin was developed for the open source software μManager to automate the phenotypic identification and photoactivation of cells, allowing ∼1.5 million individual cells to be screened in 8 h. We used this approach to screen 6,092 sgRNAs targeting 544 genes for their effects on nuclear size regulation and identified 14 bona fide hits. These results present a scalable approach to facilitate imaging-based pooled CRISPR screens. |
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| AbstractList | CRISPR (clustered regularly interspaced short palindromic repeats)-based gene inactivation provides a powerful means for linking genes to particular cellular phenotypes. CRISPR-based screening typically uses large genomic pools of single guide RNAs (sgRNAs). However, this approach is limited to phenotypes that can be enriched by chemical selection or FACS sorting. Here, we developed a microscopy-based approach, which we name optical enrichment, to select cells displaying a particular CRISPR-induced phenotype by automated imaging-based computation, mark them by photoactivation of an expressed photoactivatable fluorescent protein, and then isolate the fluorescent cells using fluorescence-activated cell sorting (FACS). A plugin was developed for the open source software μManager to automate the phenotypic identification and photoactivation of cells, allowing ∼1.5 million individual cells to be screened in 8 h. We used this approach to screen 6,092 sgRNAs targeting 544 genes for their effects on nuclear size regulation and identified 14 bona fide hits. These results present a scalable approach to facilitate imaging-based pooled CRISPR screens. Yan et al. demonstrate high-throughput screening of pooled CRISPR libraries for phenotypes detectable by microscopy. Their approach uses photoactivation of cells displaying the phenotype of interest and FACS sorting of marked cells, followed by sequencing, and facilitates discovery of genes involved in cell biological processes. CRISPR (clustered regularly interspaced short palindromic repeats)-based gene inactivation provides a powerful means for linking genes to particular cellular phenotypes. CRISPR-based screening typically uses large genomic pools of single guide RNAs (sgRNAs). However, this approach is limited to phenotypes that can be enriched by chemical selection or FACS sorting. Here, we developed a microscopy-based approach, which we name optical enrichment, to select cells displaying a particular CRISPR-induced phenotype by automated imaging-based computation, mark them by photoactivation of an expressed photoactivatable fluorescent protein, and then isolate the fluorescent cells using fluorescence-activated cell sorting (FACS). A plugin was developed for the open source software μManager to automate the phenotypic identification and photoactivation of cells, allowing ∼1.5 million individual cells to be screened in 8 h. We used this approach to screen 6,092 sgRNAs targeting 544 genes for their effects on nuclear size regulation and identified 14 bona fide hits. These results present a scalable approach to facilitate imaging-based pooled CRISPR screens. CRISPR (clustered regularly interspaced short palindromic repeats)-based gene inactivation provides a powerful means for linking genes to particular cellular phenotypes. CRISPR-based screening typically uses large genomic pools of single guide RNAs (sgRNAs). However, this approach is limited to phenotypes that can be enriched by chemical selection or FACS sorting. Here, we developed a microscopy-based approach, which we name optical enrichment, to select cells displaying a particular CRISPR-induced phenotype by automated imaging-based computation, mark them by photoactivation of an expressed photoactivatable fluorescent protein, and then isolate the fluorescent cells using fluorescence-activated cell sorting (FACS). A plugin was developed for the open source software μManager to automate the phenotypic identification and photoactivation of cells, allowing ∼1.5 million individual cells to be screened in 8 h. We used this approach to screen 6,092 sgRNAs targeting 544 genes for their effects on nuclear size regulation and identified 14 bona fide hits. These results present a scalable approach to facilitate imaging-based pooled CRISPR screens.CRISPR (clustered regularly interspaced short palindromic repeats)-based gene inactivation provides a powerful means for linking genes to particular cellular phenotypes. CRISPR-based screening typically uses large genomic pools of single guide RNAs (sgRNAs). However, this approach is limited to phenotypes that can be enriched by chemical selection or FACS sorting. Here, we developed a microscopy-based approach, which we name optical enrichment, to select cells displaying a particular CRISPR-induced phenotype by automated imaging-based computation, mark them by photoactivation of an expressed photoactivatable fluorescent protein, and then isolate the fluorescent cells using fluorescence-activated cell sorting (FACS). A plugin was developed for the open source software μManager to automate the phenotypic identification and photoactivation of cells, allowing ∼1.5 million individual cells to be screened in 8 h. We used this approach to screen 6,092 sgRNAs targeting 544 genes for their effects on nuclear size regulation and identified 14 bona fide hits. These results present a scalable approach to facilitate imaging-based pooled CRISPR screens. |
| Author | Vale, Ronald D. Jost, Marco Ribeiro, Susana A. Stuurman, Nico Horlbeck, Max A. Liem, Christina R. Weissman, Jonathan S. Yan, Xiaowei Tanenbaum, Marvin E. |
| AuthorAffiliation | 2 Cairn Biosciences, Inc., San Francisco, CA 4 Boston Children's Hospital, Boston, MA 6 Whitehead Institute and Department of Biology, MIT, Cambridge, MA 1 Department of Cellular and Molecular Pharmacology, Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, CA 7 Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 5 University of California, San Diego, San Diego, CA 3 Oncode Institute, Hubrecht Institute-KNAW and University Medical Center Utrecht, Utrecht, Netherlands |
| AuthorAffiliation_xml | – name: 5 University of California, San Diego, San Diego, CA – name: 6 Whitehead Institute and Department of Biology, MIT, Cambridge, MA – name: 7 Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA – name: 3 Oncode Institute, Hubrecht Institute-KNAW and University Medical Center Utrecht, Utrecht, Netherlands – name: 2 Cairn Biosciences, Inc., San Francisco, CA – name: 1 Department of Cellular and Molecular Pharmacology, Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, CA – name: 4 Boston Children's Hospital, Boston, MA |
| Author_xml | – sequence: 1 givenname: Xiaowei orcidid: 0000-0002-4846-8812 surname: Yan fullname: Yan, Xiaowei – sequence: 2 givenname: Nico orcidid: 0000-0002-6179-8613 surname: Stuurman fullname: Stuurman, Nico – sequence: 3 givenname: Susana A. orcidid: 0000-0003-3807-7540 surname: Ribeiro fullname: Ribeiro, Susana A. – sequence: 4 givenname: Marvin E. surname: Tanenbaum fullname: Tanenbaum, Marvin E. – sequence: 5 givenname: Max A. orcidid: 0000-0002-3875-871X surname: Horlbeck fullname: Horlbeck, Max A. – sequence: 6 givenname: Christina R. orcidid: 0000-0003-2744-6312 surname: Liem fullname: Liem, Christina R. – sequence: 7 givenname: Marco orcidid: 0000-0002-1369-4908 surname: Jost fullname: Jost, Marco – sequence: 8 givenname: Jonathan S. surname: Weissman fullname: Weissman, Jonathan S. – sequence: 9 givenname: Ronald D. orcidid: 0000-0003-3460-2758 surname: Vale fullname: Vale, Ronald D. |
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| Cites_doi | 10.1038/onc.2010.232 10.1089/scd.2014.0129 10.1016/j.cell.2018.09.022 10.1080/19491034.2016.1162933 10.1016/j.cell.2018.11.022 10.15252/msb.20209442 10.1016/j.tibtech.2018.08.002 10.1016/j.cell.2019.09.016 10.1038/nbt.3659 10.3389/fonc.2019.00516 10.1073/pnas.1903808116 10.4161/cc.26671 10.1038/s41592-018-0111-2 10.1039/C4SC03676J 10.1242/jcs.037333 10.1038/nrm3474 10.1016/j.cell.2014.09.029 10.1126/science.1247005 10.1016/j.cell.2014.05.010 10.1091/mbc.e13-07-0421 10.1247/csf.26.653 10.1038/s41589-018-0004-9 10.1101/gad.1495007 10.1038/s41592-020-0826-8 10.1016/j.cell.2015.06.059 10.1242/jcs.035360 10.3390/ijms18071562 10.1101/gad.1860310 10.1128/MCB.00908-07 10.1101/gad.1687508 10.1016/j.cell.2016.11.039 10.1038/nmeth.4177 10.1080/15384101.2016.1216928 10.1007/s12272-018-1029-z 10.1016/j.cub.2009.12.032 10.1101/2020.07.02.184390 10.1080/15384101.2016.1261765 10.15252/msb.20178064 10.1073/pnas.1307002110 10.7554/eLife.12902 10.14440/jbm.2014.36 10.7554/eLife.19760 10.1126/science.1246981 10.1128/MCB.01834-08 10.1016/j.cell.2016.11.038 10.1016/j.tcb.2012.11.004 10.1007/s00294-019-00999-3 10.3390/genes9120629 10.1101/298349 |
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| References | Cantwell (2023072812143826200_bib4) 2019; 65 Hsu (2023072812143826200_bib19) 2014; 157 Webster (2023072812143826200_bib44) 2009; 122 Kampmann (2023072812143826200_bib22) 2013; 110 Bower (2023072812143826200_bib3) 2010; 29 Chien (2023072812143826200_bib6) 2015; 6 Kweon (2023072812143826200_bib24) 2018; 41 Wheeler (2023072812143826200_bib45) 2020; 17 Parnas (2023072812143826200_bib28) 2015; 162 Jaitin (2023072812143826200_bib21) 2016; 167 Wroblewska (2023072812143826200_bib46) 2018; 175 Adamson (2023072812143826200_bib1) 2018 Horlbeck (2023072812143826200_bib18) 2016; 5 Sokolova (2023072812143826200_bib38) 2017; 16 Dixit (2023072812143826200_bib9) 2016; 167 Wang (2023072812143826200_bib43) 2014; 343 Rapley (2023072812143826200_bib31) 2008; 121 Yoshida (2023072812143826200_bib48) 2016; 15 Ito (2023072812143826200_bib20) 2010; 20 Sansam (2023072812143826200_bib33) 2010; 24 Kanfer (2023072812143826200_bib23) 2020 Edens (2023072812143826200_bib12) 2013; 23 Sivakumar (2023072812143826200_bib36) 2014; 25 Hasle (2023072812143826200_bib17) 2020; 16 Gassmann (2023072812143826200_bib14) 2008; 22 Gilbert (2023072812143826200_bib15) 2014; 159 Ounkomol (2023072812143826200_bib27) 2018; 15 Edelstein (2023072812143826200_bib11) 2014; 1 Yu (2023072812143826200_bib49) 2019; 9 Terada (2023072812143826200_bib40) 2001; 26 Barrangou (2023072812143826200_bib2) 2016; 34 Carmena (2023072812143826200_bib5) 2012; 13 Sivakumar (2023072812143826200_bib37) 2016; 5 Verschuren (2023072812143826200_bib41) 2007; 27 Machida (2023072812143826200_bib25) 2007; 21 Pawłowska (2023072812143826200_bib29) 2017; 18 Datlinger (2023072812143826200_bib7) 2017; 14 Rubin (2023072812143826200_bib32) 2019; 176 Duxin (2023072812143826200_bib10) 2009; 29 Wang (2023072812143826200_bib42) 2019; 116 Piatkevich (2023072812143826200_bib30) 2018; 14 Feldman (2023072812143826200_bib13) 2019; 179 Sullivan (2023072812143826200_bib39) 2018; 9 Yoon (2023072812143826200_bib47) 2014; 23 Schuster (2023072812143826200_bib34) 2019; 37 de Groot (2023072812143826200_bib8) 2018; 14 Mukherjee (2023072812143826200_bib26) 2016; 7 Hashizume (2023072812143826200_bib16) 2013; 12 Shalem (2023072812143826200_bib35) 2014; 343 |
| References_xml | – volume: 29 start-page: 4787 year: 2010 ident: 2023072812143826200_bib3 article-title: Topoisomerase IIalpha maintains genomic stability through decatenation G(2) checkpoint signaling publication-title: Oncogene. doi: 10.1038/onc.2010.232 – volume: 23 start-page: 2700 year: 2014 ident: 2023072812143826200_bib47 article-title: Rad51 regulates cell cycle progression by preserving G2/M transition in mouse embryonic stem cells publication-title: Stem Cells Dev. doi: 10.1089/scd.2014.0129 – volume: 175 start-page: 1141 year: 2018 ident: 2023072812143826200_bib46 article-title: Protein Barcodes Enable High-Dimensional Single-Cell CRISPR Screens publication-title: Cell. doi: 10.1016/j.cell.2018.09.022 – volume: 7 start-page: 167 year: 2016 ident: 2023072812143826200_bib26 article-title: Recent advances in understanding nuclear size and shape publication-title: Nucleus. doi: 10.1080/19491034.2016.1162933 – volume: 176 start-page: 361 year: 2019 ident: 2023072812143826200_bib32 article-title: Coupled Single-Cell CRISPR Screening and Epigenomic Profiling Reveals Causal Gene Regulatory Networks publication-title: Cell. doi: 10.1016/j.cell.2018.11.022 – volume: 16 year: 2020 ident: 2023072812143826200_bib17 article-title: High-throughput, microscope-based sorting to dissect cellular heterogeneity publication-title: Mol. Syst. Biol. doi: 10.15252/msb.20209442 – volume: 37 start-page: 38 year: 2019 ident: 2023072812143826200_bib34 article-title: RNAi/CRISPR Screens: from a Pool to a Valid Hit publication-title: Trends Biotechnol. doi: 10.1016/j.tibtech.2018.08.002 – volume: 179 start-page: 787 year: 2019 ident: 2023072812143826200_bib13 article-title: Optical Pooled Screens in Human Cells publication-title: Cell. doi: 10.1016/j.cell.2019.09.016 – volume: 34 start-page: 933 year: 2016 ident: 2023072812143826200_bib2 article-title: Applications of CRISPR technologies in research and beyond publication-title: Nat. Biotechnol. doi: 10.1038/nbt.3659 – volume: 9 start-page: 516 year: 2019 ident: 2023072812143826200_bib49 article-title: TICRR Contributes to Tumorigenesis Through Accelerating DNA Replication in Cancers publication-title: Front. Oncol. doi: 10.3389/fonc.2019.00516 – volume: 116 start-page: 10842 year: 2019 ident: 2023072812143826200_bib42 article-title: Imaging-based pooled CRISPR screening reveals regulators of lncRNA localization publication-title: Proc. Natl. Acad. Sci. USA. doi: 10.1073/pnas.1903808116 – volume: 12 start-page: 3804 year: 2013 ident: 2023072812143826200_bib16 article-title: Nucleoporin Nup62 maintains centrosome homeostasis publication-title: Cell Cycle. doi: 10.4161/cc.26671 – volume: 15 start-page: 917 year: 2018 ident: 2023072812143826200_bib27 article-title: Label-free prediction of three-dimensional fluorescence images from transmitted-light microscopy publication-title: Nat. Methods. doi: 10.1038/s41592-018-0111-2 – volume: 6 start-page: 1701 year: 2015 ident: 2023072812143826200_bib6 article-title: Photostick: a method for selective isolation of target cells from culture publication-title: Chem. Sci. (Camb.). doi: 10.1039/C4SC03676J – volume: 122 start-page: 1477 year: 2009 ident: 2023072812143826200_bib44 article-title: Sizing up the nucleus: nuclear shape, size and nuclear-envelope assembly publication-title: J. Cell Sci. doi: 10.1242/jcs.037333 – volume: 13 start-page: 789 year: 2012 ident: 2023072812143826200_bib5 article-title: The chromosomal passenger complex (CPC): from easy rider to the godfather of mitosis publication-title: Nat. Rev. Mol. Cell Biol. doi: 10.1038/nrm3474 – volume: 159 start-page: 647 year: 2014 ident: 2023072812143826200_bib15 article-title: Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation publication-title: Cell. doi: 10.1016/j.cell.2014.09.029 – volume: 343 start-page: 84 year: 2014 ident: 2023072812143826200_bib35 article-title: Genome-scale CRISPR-Cas9 knockout screening in human cells publication-title: Science. doi: 10.1126/science.1247005 – volume: 157 start-page: 1262 year: 2014 ident: 2023072812143826200_bib19 article-title: Development and applications of CRISPR-Cas9 for genome engineering publication-title: Cell. doi: 10.1016/j.cell.2014.05.010 – volume: 25 start-page: 594 year: 2014 ident: 2023072812143826200_bib36 article-title: The spindle and kinetochore-associated (Ska) complex enhances binding of the anaphase-promoting complex/cyclosome (APC/C) to chromosomes and promotes mitotic exit publication-title: Mol. Biol. Cell. doi: 10.1091/mbc.e13-07-0421 – volume: 26 start-page: 653 year: 2001 ident: 2023072812143826200_bib40 article-title: Role of chromosomal passenger complex in chromosome segregation and cytokinesis publication-title: Cell Struct. Funct. doi: 10.1247/csf.26.653 – volume: 14 start-page: 352 year: 2018 ident: 2023072812143826200_bib30 article-title: A robotic multidimensional directed evolution approach applied to fluorescent voltage reporters publication-title: Nat. Chem. Biol. doi: 10.1038/s41589-018-0004-9 – volume: 21 start-page: 184 year: 2007 ident: 2023072812143826200_bib25 article-title: The APC/C inhibitor, Emi1, is essential for prevention of rereplication publication-title: Genes Dev. doi: 10.1101/gad.1495007 – volume: 17 start-page: 636 year: 2020 ident: 2023072812143826200_bib45 article-title: Pooled CRISPR screens with imaging on microraft arrays reveals stress granule-regulatory factors publication-title: Nat. Methods. doi: 10.1038/s41592-020-0826-8 – volume: 162 start-page: 675 year: 2015 ident: 2023072812143826200_bib28 article-title: A Genome-wide CRISPR Screen in Primary Immune Cells to Dissect Regulatory Networks publication-title: Cell. doi: 10.1016/j.cell.2015.06.059 – volume: 121 start-page: 3912 year: 2008 ident: 2023072812143826200_bib31 article-title: The NIMA-family kinase Nek6 phosphorylates the kinesin Eg5 at a novel site necessary for mitotic spindle formation publication-title: J. Cell Sci. doi: 10.1242/jcs.035360 – volume: 18 start-page: 1562 year: 2017 ident: 2023072812143826200_bib29 article-title: DNA2-An Important Player in DNA Damage Response or Just Another DNA Maintenance Protein? publication-title: Int. J. Mol. Sci. doi: 10.3390/ijms18071562 – volume: 24 start-page: 183 year: 2010 ident: 2023072812143826200_bib33 article-title: A vertebrate gene, ticrr, is an essential checkpoint and replication regulator publication-title: Genes Dev. doi: 10.1101/gad.1860310 – volume: 27 start-page: 7955 year: 2007 ident: 2023072812143826200_bib41 article-title: Loss of Emi1-dependent anaphase-promoting complex/cyclosome inhibition deregulates E2F target expression and elicits DNA damage-induced senescence publication-title: Mol. Cell. Biol. doi: 10.1128/MCB.00908-07 – volume: 22 start-page: 2385 year: 2008 ident: 2023072812143826200_bib14 article-title: A new mechanism controlling kinetochore-microtubule interactions revealed by comparison of two dynein-targeting components: SPDL-1 and the Rod/Zwilch/Zw10 complex publication-title: Genes Dev. doi: 10.1101/gad.1687508 – volume: 167 start-page: 1883 year: 2016 ident: 2023072812143826200_bib21 article-title: Dissecting Immune Circuits by Linking CRISPR-Pooled Screens with Single-Cell RNA-Seq publication-title: Cell. doi: 10.1016/j.cell.2016.11.039 – volume: 14 start-page: 297 year: 2017 ident: 2023072812143826200_bib7 article-title: Pooled CRISPR screening with single-cell transcriptome readout publication-title: Nat. Methods. doi: 10.1038/nmeth.4177 – volume: 15 start-page: 3151 year: 2016 ident: 2023072812143826200_bib48 article-title: Mechanisms behind Topoisomerase II SUMOylation in chromosome segregation publication-title: Cell Cycle. doi: 10.1080/15384101.2016.1216928 – volume: 41 start-page: 875 year: 2018 ident: 2023072812143826200_bib24 article-title: High-throughput genetic screens using CRISPR-Cas9 system publication-title: Arch. Pharm. Res. doi: 10.1007/s12272-018-1029-z – volume: 20 start-page: 333 year: 2010 ident: 2023072812143826200_bib20 article-title: Cell-nonautonomous regulation of C. elegans germ cell death by kri-1 publication-title: Curr. Biol. doi: 10.1016/j.cub.2009.12.032 – year: 2020 ident: 2023072812143826200_bib23 doi: 10.1101/2020.07.02.184390 – volume: 16 start-page: 189 year: 2017 ident: 2023072812143826200_bib38 article-title: Genome-wide screen of cell-cycle regulators in normal and tumor cells identifies a differential response to nucleosome depletion publication-title: Cell Cycle. doi: 10.1080/15384101.2016.1261765 – volume: 14 year: 2018 ident: 2023072812143826200_bib8 article-title: Large-scale image-based profiling of single-cell phenotypes in arrayed CRISPR-Cas9 gene perturbation screens publication-title: Mol. Syst. Biol. doi: 10.15252/msb.20178064 – volume: 110 start-page: E2317 year: 2013 ident: 2023072812143826200_bib22 article-title: Integrated platform for genome-wide screening and construction of high-density genetic interaction maps in mammalian cells publication-title: Proc. Natl. Acad. Sci. USA. doi: 10.1073/pnas.1307002110 – volume: 5 year: 2016 ident: 2023072812143826200_bib37 article-title: The human SKA complex drives the metaphase-anaphase cell cycle transition by recruiting protein phosphatase 1 to kinetochores publication-title: eLife. doi: 10.7554/eLife.12902 – volume: 1 start-page: 10 year: 2014 ident: 2023072812143826200_bib11 article-title: Advanced methods of microscope control using μManager software publication-title: J. Biol. Methods. doi: 10.14440/jbm.2014.36 – volume: 5 year: 2016 ident: 2023072812143826200_bib18 article-title: Compact and highly active next-generation libraries for CRISPR-mediated gene repression and activation publication-title: eLife. doi: 10.7554/eLife.19760 – volume: 343 start-page: 80 year: 2014 ident: 2023072812143826200_bib43 article-title: Genetic screens in human cells using the CRISPR-Cas9 system publication-title: Science. doi: 10.1126/science.1246981 – volume: 29 start-page: 4274 year: 2009 ident: 2023072812143826200_bib10 article-title: Human Dna2 is a nuclear and mitochondrial DNA maintenance protein publication-title: Mol. Cell. Biol. doi: 10.1128/MCB.01834-08 – volume: 167 start-page: 1853 year: 2016 ident: 2023072812143826200_bib9 article-title: Perturb-Seq: Dissecting Molecular Circuits with Scalable Single-Cell RNA Profiling of Pooled Genetic Screens publication-title: Cell. doi: 10.1016/j.cell.2016.11.038 – volume: 23 start-page: 151 year: 2013 ident: 2023072812143826200_bib12 article-title: Nuclear size regulation: from single cells to development and disease publication-title: Trends Cell Biol. doi: 10.1016/j.tcb.2012.11.004 – volume: 65 start-page: 1281 year: 2019 ident: 2023072812143826200_bib4 article-title: Unravelling nuclear size control publication-title: Curr. Genet. doi: 10.1007/s00294-019-00999-3 – volume: 9 start-page: 629 year: 2018 ident: 2023072812143826200_bib39 article-title: RAD-ical New Insights into RAD51 Regulation publication-title: Genes (Basel). doi: 10.3390/genes9120629 – year: 2018 ident: 2023072812143826200_bib1 doi: 10.1101/298349 |
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| Snippet | CRISPR (clustered regularly interspaced short palindromic repeats)-based gene inactivation provides a powerful means for linking genes to particular cellular... Yan et al. demonstrate high-throughput screening of pooled CRISPR libraries for phenotypes detectable by microscopy. Their approach uses photoactivation of... |
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| SubjectTerms | Automation Cell Cycle and Division Cell Line Cell Nucleus - genetics Cell Nucleus Size - genetics CRISPR CRISPR-Cas Systems - genetics Flow Cytometry Fluorescence Genes Genetic Testing Green Fluorescent Proteins - metabolism Humans Imaging Imaging, Three-Dimensional Inactivation Mammalian cells Optics and Photonics Phenotype Phenotypes Photoactivation Technology |
| Title | High-content imaging-based pooled CRISPR screens in mammalian cells |
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