{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T19:02:33Z","timestamp":1785438153700,"version":"3.56.0"},"posted":{"date-parts":[[2026,7,27]]},"group-title":"Cell Biology","reference-count":61,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T00:00:00Z","timestamp":1785110400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"award":["R35GM126930"],"award-info":[{"award-number":["R35GM126930"]}],"id":[{"id":"https:\/\/ror.org\/04q48ey07","id-type":"ROR","asserted-by":"publisher"}]},{"award":["Cell Biology @ Scale"],"award-info":[{"award-number":["Cell Biology @ Scale"]}],"id":[{"id":"https:\/\/ror.org\/02qenvm24","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000065","name":"National Institute of Neurological Disorders and Stroke","doi-asserted-by":"crossref","award":["RM1NS133601"],"award-info":[{"award-number":["RM1NS133601"]}],"id":[{"id":"10.13039\/100000065","id-type":"DOI","asserted-by":"crossref"}]},{"award":["R61CA278536"],"award-info":[{"award-number":["R61CA278536"]}],"id":[{"id":"https:\/\/ror.org\/040gcmg81","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"crossref","award":["P01AI120943"],"award-info":[{"award-number":["P01AI120943"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]},{"award":["Graduate Research Fellowship"],"award-info":[{"award-number":["Graduate Research Fellowship"]}],"id":[{"id":"https:\/\/ror.org\/021nxhr62","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,7,27]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Cellular adhesion is critical for tissue organization and integrity, but the full complement of proteins required for proper adhesion remains unresolved. Here, we define the requirements for cell-substrate adhesion in cultured human cells using orthogonal, large-scale functional genetic approaches. Using mechanical assays to test the maintenance (\u201cshake-off\u201d) or formation of adhesion and parallel large-scale assays of cell morphology, we identify dozens of gene targets with roles in adhesion. Our analyses reveal dynamic requirements for adhesion across timepoints and cell lines. We additionally conduct targeted downstream mechanistic analyses to resolve the molecular basis for altered adhesion. Collectively, we identify established and uncharacterized regulators of adhesion, including genes involved in mitosis, focal adhesions, actin regulation, and membrane trafficking. Unexpectedly, we find that cells that fail cytokinesis display impaired adhesion, with strongly altered actin organization and nuclear dynamics. Together, this work provides a comprehensive view of the genetic requirements for cell-substrate adhesion.<\/jats:p>","DOI":"10.64898\/2026.07.24.740569","type":"posted-content","created":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T13:20:39Z","timestamp":1785158439000},"source":"Crossref","is-referenced-by-count":0,"title":["Systematic functional genetic analysis of cell-substrate adhesion"],"prefix":"10.64898","author":[{"given":"Kaitlyn","family":"Manzer","sequence":"first","affiliation":[{"name":"Whitehead Institute for Biomedical Research, Cambridge, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kuan-Chung","family":"Su","sequence":"additional","affiliation":[{"name":"Whitehead Institute for Biomedical Research, Cambridge, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matteo","family":"Di Bernardo","sequence":"additional","affiliation":[{"name":"Whitehead Institute for Biomedical Research, Cambridge, United States"},{"name":"Computational and Systems Biology Program, Massachusetts Institute of Technology, Cambridge, MA 02142"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heather R.","family":"Keys","sequence":"additional","affiliation":[{"name":"Whitehead Institute for Biomedical Research, Cambridge, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Russell","family":"Walton","sequence":"additional","affiliation":[{"name":"Broad Institute of MIT and Harvard; 415 Main St., Cambridge, MA 02142, USA"},{"name":"Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02142"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brittania","family":"Moodie","sequence":"additional","affiliation":[{"name":"Whitehead Institute for Biomedical Research, Cambridge, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bingbing","family":"Yuan","sequence":"additional","affiliation":[{"name":"Whitehead Institute for Biomedical Research, Cambridge, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4889-8783","authenticated-orcid":false,"given":"Paul C.","family":"Blainey","sequence":"additional","affiliation":[{"name":"Broad Institute of MIT and Harvard; 415 Main St., Cambridge, MA 02142, USA"},{"name":"Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02142"},{"name":"Koch Institute for Integrative Cancer Research at MIT, Cambridge, MA 02142"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Iain","family":"Cheeseman","sequence":"additional","affiliation":[{"name":"Whitehead Institute for Biomedical Research, Cambridge, United States"},{"name":"Department of Biology, Massachusetts Institute of Technology, Cambridge, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026072821301071000_2026.07.24.740569v1.1","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00036.2018"},{"key":"2026072821301071000_2026.07.24.740569v1.2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0008560"},{"key":"2026072821301071000_2026.07.24.740569v1.3","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(95)90142-6"},{"key":"2026072821301071000_2026.07.24.740569v1.4","doi-asserted-by":"publisher","DOI":"10.1126\/science.279.5349.349"},{"key":"2026072821301071000_2026.07.24.740569v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.tcb.2016.07.004"},{"key":"2026072821301071000_2026.07.24.740569v1.6","doi-asserted-by":"publisher","DOI":"10.1038\/s41580-024-00777-1"},{"key":"2026072821301071000_2026.07.24.740569v1.7","doi-asserted-by":"crossref","first-page":"e2315707121","DOI":"10.1073\/pnas.2315707121","article-title":"KCTD10 regulates brain development by destabilizing brain disorder-associated protein KCTD13","volume":"121","year":"2024","journal-title":"Proc Natl Acad Sci U S A"},{"key":"2026072821301071000_2026.07.24.740569v1.8","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkaa669"},{"key":"2026072821301071000_2026.07.24.740569v1.9","unstructured":"DepMap, B. 2025. DepMap Public 25Q3."},{"key":"2026072821301071000_2026.07.24.740569v1.10","doi-asserted-by":"crossref","unstructured":"Di Bernardo, M. , R.S. Kern , A. Mallar , A. Nutter-Upham , P.C. Blainey , and I. Cheeseman . 2025. Brieflow: An Integrated Computational Pipeline for High-Throughput Analysis of Optical Pooled Screening Data. bioRxiv.","DOI":"10.1101\/2025.05.26.656231"},{"key":"2026072821301071000_2026.07.24.740569v1.11","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.3437"},{"key":"2026072821301071000_2026.07.24.740569v1.12","doi-asserted-by":"crossref","unstructured":"Drepanos, L.M. , S. Srikanth , E.G. Kaplan , S.T. Shah , B.E. Velasco , S. Merzouk , and J.G. Doench . 2026. Balancing off-target and on-target considerations for optimized CRISPR-Cas9 knockout library design. Cell Genom:101190.","DOI":"10.1016\/j.xgen.2026.101190"},{"key":"2026072821301071000_2026.07.24.740569v1.13","doi-asserted-by":"publisher","DOI":"10.1038\/s41596-021-00653-8"},{"key":"2026072821301071000_2026.07.24.740569v1.14","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2019.09.016"},{"key":"2026072821301071000_2026.07.24.740569v1.15","doi-asserted-by":"crossref","first-page":"39","DOI":"10.4161\/fly.23506","article-title":"An emerging role for Cullin-3 mediated ubiquitination in sleep and circadian rhythm: insights from Drosophila","volume":"7","year":"2013","journal-title":"Fly (Austin"},{"key":"2026072821301071000_2026.07.24.740569v1.16","doi-asserted-by":"publisher","DOI":"10.1038\/nature04217"},{"key":"2026072821301071000_2026.07.24.740569v1.17","doi-asserted-by":"publisher","DOI":"10.1126\/science.aag2445"},{"key":"2026072821301071000_2026.07.24.740569v1.18","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2022.10.017"},{"key":"2026072821301071000_2026.07.24.740569v1.19","doi-asserted-by":"publisher","DOI":"10.1016\/j.gde.2007.02.011"},{"key":"2026072821301071000_2026.07.24.740569v1.20","doi-asserted-by":"publisher","DOI":"10.1038\/emboj.2013.173"},{"key":"2026072821301071000_2026.07.24.740569v1.21","doi-asserted-by":"publisher","DOI":"10.1126\/science.1096896"},{"key":"2026072821301071000_2026.07.24.740569v1.22","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.141.5.1193"},{"key":"2026072821301071000_2026.07.24.740569v1.23","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2013.02.031"},{"key":"2026072821301071000_2026.07.24.740569v1.24","doi-asserted-by":"publisher","DOI":"10.1126\/science.1061376"},{"key":"2026072821301071000_2026.07.24.740569v1.25","doi-asserted-by":"crossref","unstructured":"Haykal, M.M. , S. Rodrigues-Ferreira , and C. Nahmias . 2024. WEE1 kinase inhibition triggers severe chromosome pulverization in aneuploid cells. bioRxiv.","DOI":"10.1101\/2023.09.19.558475"},{"key":"2026072821301071000_2026.07.24.740569v1.26","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.112722"},{"key":"2026072821301071000_2026.07.24.740569v1.27","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(02)00971-6"},{"key":"2026072821301071000_2026.07.24.740569v1.28","doi-asserted-by":"crossref","first-page":"16669","DOI":"10.18632\/oncotarget.14291","article-title":"Heterogeneity in sarcoma cell lines reveals enhanced motility of tetraploid versus diploid cells","volume":"8","year":"2017","journal-title":"Oncotarget"},{"key":"2026072821301071000_2026.07.24.740569v1.29","doi-asserted-by":"crossref","unstructured":"Jemaa, M. , R. Daams , S. Charfi , F. Mertens , S.M. Huber , and R. Massoumi . 2023. Tetraploidization Increases the Motility and Invasiveness of Cancer Cells. Int J Mol Sci. 24.","DOI":"10.3390\/ijms241813926"},{"key":"2026072821301071000_2026.07.24.740569v1.30","doi-asserted-by":"crossref","unstructured":"Kirby, B. , M. Di Bernardo , I.M. Cheeseman , and P.C. Blainey . 2026. The one-week automated genome-wide optical pooled screen. bioRxiv.","DOI":"10.64898\/2026.04.15.718742"},{"key":"2026072821301071000_2026.07.24.740569v1.31","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamcr.2018.07.023"},{"key":"2026072821301071000_2026.07.24.740569v1.32","doi-asserted-by":"publisher","DOI":"10.1038\/ncb2440"},{"key":"2026072821301071000_2026.07.24.740569v1.33","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.201606055"},{"key":"2026072821301071000_2026.07.24.740569v1.34","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.135.6.1701"},{"key":"2026072821301071000_2026.07.24.740569v1.35","doi-asserted-by":"crossref","unstructured":"Legerstee, K. , and A.B. Houtsmuller . 2021. A Layered View on Focal Adhesions. Biology (Basel ). 10.","DOI":"10.3390\/biology10111189"},{"key":"2026072821301071000_2026.07.24.740569v1.36","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-014-0554-4"},{"key":"2026072821301071000_2026.07.24.740569v1.37","doi-asserted-by":"crossref","first-page":"490","DOI":"10.1038\/s41586-024-08088-3","article-title":"Nuclear release of eIF1 restricts start-codon selection during mitosis","volume":"635","year":"2024","journal-title":"Nature"},{"key":"2026072821301071000_2026.07.24.740569v1.38","doi-asserted-by":"crossref","unstructured":"Marescal, O. , and I.M. Cheeseman . 2025. 19S proteasome loss causes monopolar spindles through ubiquitin-independent KIF11 degradation. bioRxiv.","DOI":"10.1101\/2025.01.08.632038"},{"key":"2026072821301071000_2026.07.24.740569v1.39","doi-asserted-by":"publisher","DOI":"10.1016\/j.devcel.2017.01.012"},{"key":"2026072821301071000_2026.07.24.740569v1.40","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2015.10.027"},{"key":"2026072821301071000_2026.07.24.740569v1.41","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-391857-4.00009-4"},{"key":"2026072821301071000_2026.07.24.740569v1.42","doi-asserted-by":"publisher","DOI":"10.1016\/j.devcel.2004.06.006"},{"key":"2026072821301071000_2026.07.24.740569v1.43","doi-asserted-by":"publisher","DOI":"10.1111\/cas.13899"},{"key":"2026072821301071000_2026.07.24.740569v1.44","doi-asserted-by":"publisher","DOI":"10.1038\/nrm4012"},{"key":"2026072821301071000_2026.07.24.740569v1.45","doi-asserted-by":"crossref","unstructured":"Pachitariu, M. , M. Rariden , and C. Stringer . 2025. Cellpose-SAM: superhuman generalization for cellular segmentation. bioRxiv.","DOI":"10.1101\/2025.04.28.651001"},{"key":"2026072821301071000_2026.07.24.740569v1.46","doi-asserted-by":"publisher","DOI":"10.1038\/nrm2957"},{"key":"2026072821301071000_2026.07.24.740569v1.47","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkae1011"},{"key":"2026072821301071000_2026.07.24.740569v1.48","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2018488117"},{"key":"2026072821301071000_2026.07.24.740569v1.49","doi-asserted-by":"publisher","DOI":"10.1016\/j.devcel.2021.01.016"},{"key":"2026072821301071000_2026.07.24.740569v1.50","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-07901-8"},{"key":"2026072821301071000_2026.07.24.740569v1.51","doi-asserted-by":"publisher","DOI":"10.1126\/science.1247005"},{"key":"2026072821301071000_2026.07.24.740569v1.52","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M700241200"},{"key":"2026072821301071000_2026.07.24.740569v1.53","doi-asserted-by":"publisher","DOI":"10.1186\/s12859-021-04344-9"},{"key":"2026072821301071000_2026.07.24.740569v1.54","doi-asserted-by":"publisher","DOI":"10.1126\/science.1240104"},{"key":"2026072821301071000_2026.07.24.740569v1.55","doi-asserted-by":"crossref","first-page":"687","DOI":"10.3389\/fcell.2020.00687","article-title":"The Mechanics of Mitotic Cell Rounding","volume":"8","year":"2020","journal-title":"Front Cell Dev Biol"},{"key":"2026072821301071000_2026.07.24.740569v1.56","doi-asserted-by":"publisher","DOI":"10.1007\/s00109-014-1133-6"},{"key":"2026072821301071000_2026.07.24.740569v1.57","doi-asserted-by":"crossref","unstructured":"Walton, R.T. , Y. Qin , B. Kirby , J.O. Andrews , M. Taipale , B. Kang , and P.C. Blainey . 2025. CROPseq-multi: a universal solution for multiplexed perturbation in high-content pooled CRISPR screens. bioRxiv.","DOI":"10.1101\/2024.03.17.585235"},{"key":"2026072821301071000_2026.07.24.740569v1.58","doi-asserted-by":"publisher","DOI":"10.1152\/ajpcell.00377.2010"},{"key":"2026072821301071000_2026.07.24.740569v1.59","doi-asserted-by":"publisher","DOI":"10.1126\/science.aac7041"},{"key":"2026072821301071000_2026.07.24.740569v1.60","doi-asserted-by":"publisher","DOI":"10.1126\/science.1246981"},{"key":"2026072821301071000_2026.07.24.740569v1.61","doi-asserted-by":"publisher","DOI":"10.1016\/j.ceb.2011.11.001"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.07.24.740569","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T04:30:22Z","timestamp":1785299422000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.07.24.740569"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,27]]},"references-count":61,"URL":"https:\/\/doi.org\/10.64898\/2026.07.24.740569","relation":{},"subject":[],"published":{"date-parts":[[2026,7,27]]},"subtype":"preprint"}}