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Because the most notorious member of this clade,\n                  <jats:italic>Mycobacterium tuberculosis,<\/jats:italic>\n                  is a major human pathogen, research on Mycobacteriales has focused on pathogenesis, and, as a consequence, many fundamental aspects of Mycobacterial biology remain understudied. Here, we address this gap by performing a genome-wide CRISPRi chemical genomics screen using a diverse set of &gt;35 antibiotics, detergents, and other anti-microbials predominantly targeting the cell envelope of\n                  <jats:italic>Mycobacterium smegmatis,<\/jats:italic>\n                  a saprophytic model Mycobacterium. We highlight new information derived from this screen, including the identification of novel functions for previously uncharacterized conserved and essential genes (in mycolic acid and arabinogalactan synthesis), the discovery of a new drug scaffold\/protein target pair, and insights into the mechanism of action of two commonly used antibiotics. These data are also a valuable resource for the mycobacterial research community, as they provide thousands of novel phenotypes for uncharacterized genes and meaningful phenotypic correlations between annotated and uncharacterized genes.\n                <\/jats:p>","DOI":"10.1101\/2025.11.14.688347","type":"posted-content","created":{"date-parts":[[2025,11,16]],"date-time":"2025-11-16T03:50:12Z","timestamp":1763265012000},"source":"Crossref","is-referenced-by-count":5,"title":["The phenotypic landscape of the mycobacterial cell"],"prefix":"10.64898","author":[{"given":"Nadia","family":"Herrera","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94158, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Horia","family":"Todor","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94158, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lili M.","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94158, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hannah N.","family":"Burkhart","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94158, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Evan","family":"Billings","sequence":"additional","affiliation":[{"name":"Department of Physiology and Cellular Biophysics, Columbia University Irving Medical Center, New York, 10032, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Allison","family":"Fay","sequence":"additional","affiliation":[{"name":"Immunology Program, Sloan-Kettering Institute, New York, NY 10065, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Theodore C.","family":"Warner","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"So Young","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natalie Y.","family":"Sayegh","sequence":"additional","affiliation":[{"name":"Laboratory of Host-Pathogen Biology, The Rockefeller University, New York, NY 10065, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Bosch","sequence":"additional","affiliation":[{"name":"Laboratory of Host-Pathogen Biology, The Rockefeller University, New York, NY 10065, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94158, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laura L.","family":"Kiessling","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States"},{"name":"The Broad Institute of MIT and Harvard; Cambridge, MA, USA"},{"name":"Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology; Cambridge, MA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael S.","family":"Glickman","sequence":"additional","affiliation":[{"name":"Immunology Program, Sloan-Kettering Institute, New York, NY 10065, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Filippo","family":"Mancia","sequence":"additional","affiliation":[{"name":"Department of Physiology and Cellular Biophysics, Columbia University Irving Medical Center, New York, 10032, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeremy M.","family":"Rock","sequence":"additional","affiliation":[{"name":"Laboratory of Host-Pathogen Biology, The Rockefeller University, New York, NY 10065, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carol A.","family":"Gross","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94158, USA"},{"name":"Department of Cell and Tissue Biology, University of California, San Francisco, San Francisco, CA 94158, USA"},{"name":"California Institute of Quantitative Biology, University of California, San Francisco, San Francisco 94158, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026020503160426000_2025.11.14.688347v3.1","doi-asserted-by":"crossref","first-page":"1154","DOI":"10.1038\/s41396-020-0598-2","article-title":"Synergism between Corynebacterium and Streptococcus sanguinis reveals new interactions between oral commensals","volume":"14","year":"2020","journal-title":"ISME J"},{"key":"2026020503160426000_2025.11.14.688347v3.2","unstructured":"Global tuberculosis report 2024 (2024). https:\/\/www.who.int\/publications\/i\/item\/9789240101531."},{"key":"2026020503160426000_2025.11.14.688347v3.3","doi-asserted-by":"crossref","first-page":"1157","DOI":"10.1038\/s41564-024-01683-0","article-title":"Rethinking the burden of latent tuberculosis to reprioritize research","volume":"9","year":"2024","journal-title":"Nat. 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