biostudies-literatureLissner REuropean Research CouncilDutch Research Council (NWO)ZonMwe0211925https://www.ebi.ac.uk/biostudies/studies/S-EPMC12802301biostudies-literatureUnknown17(1)Accounting for more deaths than any other bacterial species, <i>Mycobacterium tuberculosis</i> (Mtb) represents a critical threat to public health worldwide. A key factor contributing to Mtb's virulence is its unique cell envelope, which acts as a protective barrier. Among the components of this envelope, lipoproteins represent a critical but understudied group of proteins. In this study, we focused on 79 conserved putative lipoproteins, shared between Mtb and the closely related <i>M. marinum</i>. Leveraging the CRISPR/Cas9 gene editing system for Mycobacteria, we generated frameshift mutations, targeting one conserved lipoprotein-coding gene at a time. We identified two mutants, <i>lpqZ</i> and <i>fecB</i>, that exhibited increased susceptibility to all tested antibiotics, suggesting crimBioA new role for lipoproteins LpqZ and FecB in orchestrating mycobacterial cell envelope biogenesis.PMC12802301XL21.00609120012010040OCENW.KLEIN.439832721Lissner RBenedict STSpeer AFranklin AKnol JNejentsev SKuijl CBitter WCharitou VMoynihan PJJimenez CfalseA new role for lipoproteins LpqZ and FecB in orchestrating mycobacterial cell envelope biogenesis.Accounting for more deaths than any other bacterial species, <i>Mycobacterium tuberculosis</i> (Mtb) represents a critical threat to public health worldwide. A key factor contributing to Mtb's virulence is its unique cell envelope, which acts as a protective barrier. Among the components of this envelope, lipoproteins represent a critical but understudied group of proteins. In this study, we focused on 79 conserved putative lipoproteins, shared between Mtb and the closely related <i>M. marinum</i>. Leveraging the CRISPR/Cas9 gene editing system for Mycobacteria, we generated frameshift mutations, targeting one conserved lipoprotein-coding gene at a time. We identified two mutants, <i>lpqZ</i> and <i>fecB</i>, that exhibited increased susceptibility to all tested antibiotics, suggesting cri2026-01-01T00:00:00Z2026 Jan2026-06-06T13:17:34.374Z2026-05-31T03:07:14.625ZS-EPMC128023014138496910.1128/mbio.02119-25