{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T01:16:37Z","timestamp":1788225397932,"version":"build-2803163510"},"reference-count":64,"publisher":"American Society for Microbiology","issue":"7","license":[{"start":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T00:00:00Z","timestamp":1783382400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T00:00:00Z","timestamp":1783382400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2026,7,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      The global rise of convergent carbapenem-resistant and hypervirulent\n                      <jats:italic toggle=\"yes\">Klebsiella pneumoniae<\/jats:italic>\n                      (CR-hvKp) represents a major clinical challenge, yet the role of virulence plasmids (pVirs) in shaping bacterial physiology and pathogenicity remains incompletely understood. Using a CRISPR-Cas9-based curing system, we precisely eliminated pVirs from two clinical CR-hvKp strains, sequence type 23 (ST23)-KL1 (\n                      <jats:italic toggle=\"yes\">bla<\/jats:italic>\n                      <jats:sub>NDM-1<\/jats:sub>\n                      ) and ST11-KL64 (\n                      <jats:italic toggle=\"yes\">bla<\/jats:italic>\n                      <jats:sub>KPC-2<\/jats:sub>\n                      ), with distinct genetic backgrounds. Loss of pVir conferred fitness advantages\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      , reduced capsule production and hypermucoviscosity, and promoted biofilm formation, while markedly attenuating virulence in murine sepsis models. Despite this reduction, the pVir-cured ST23-KL1 strain retained higher virulence than the pVir-cured ST11-KL64 strain, underscoring the contribution of chromosomal background to pathogenic potential. Transcriptomic profiling revealed both shared and strain-specific transcriptional responses to pVir deletion, with broader perturbations observed in the ST11-KL64 strain. pVir removal had limited effects on antibiotic minimum inhibitory concentrations. Complementation experiments further demonstrated differential regulatory roles of the\n                      <jats:italic toggle=\"yes\">rmpADC<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">rmpA2D2<\/jats:italic>\n                      operons in capsule expression and hypermucoviscosity across the two strains. Together, these findings establish pVirs as central determinants of CR-hvKp virulence and highlight complex host-plasmid interactions that influence bacterial adaptation and pathogenicity.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        The emergence of carbapenem-resistant and hypervirulent\n                        <jats:italic toggle=\"yes\">Klebsiella pneumoniae<\/jats:italic>\n                        (CR-hvKp) poses a critical threat to global health, yet the contribution of virulence plasmids (pVirs) to bacterial fitness and pathogenicity remains poorly defined. By employing a CRISPR-Cas9-based curing strategy, we dissected the role of pVirs in two genetically distinct CR-hvKp strains and uncovered their multifaceted impact on capsule production, hypermucoviscosity, biofilm formation, and virulence. Our findings reveal that pVir loss confers fitness advantages\n                        <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                        while attenuating virulence\n                        <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                        , with strain-specific transcriptional responses and differential regulation by\n                        <jats:italic toggle=\"yes\">rmp<\/jats:italic>\n                        operons. These results underscore the complex interplay between plasmid-encoded and chromosomal determinants in shaping CR-hvKp pathogenicity and adaptation, offering mechanistic insights that may inform future therapeutic strategies targeting plasmid-mediated virulence.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.04174-25","type":"journal-article","created":{"date-parts":[[2026,6,9]],"date-time":"2026-06-09T13:03:03Z","timestamp":1781010183000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Distinct and shared impacts of virulence plasmids on the phenotype and transcriptome in convergent carbapenem-resistant and hypervirulent\n                    <i>Klebsiella pneumoniae<\/i>"],"prefix":"10.1128","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2714-3998","authenticated-orcid":true,"given":"Mingju","family":"Hao","sequence":"first","affiliation":[{"name":"Department of Clinical Laboratory Medicine, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital","place":["Jinan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaodi","family":"Cui","sequence":"additional","affiliation":[{"name":"Wen-shang County People\u2019s Hospital","place":["Wenshang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liya","family":"Feng","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Medicine, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital","place":["Jinan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ke","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Medicine, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital","place":["Jinan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohong","family":"Shi","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Medicine, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital","place":["Jinan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tengfei","family":"Long","sequence":"additional","affiliation":[{"name":"Division of Clinical and Translational Therapeutics, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo","place":["Buffalo, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8955-359X","authenticated-orcid":false,"given":"Sarah E.","family":"Rowe","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of North Carolina","place":["Chapel Hill, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9940-7529","authenticated-orcid":true,"given":"Yi-Tsung","family":"Lin","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03ymy8z76","id-type":"ROR","asserted-by":"publisher"}],"name":"Division of Infectious Diseases, Department of Medicine, Taipei Veterans General Hospital","place":["Taipei, Taiwan"]},{"id":[{"id":"https:\/\/ror.org\/00se2k293","id-type":"ROR","asserted-by":"publisher"}],"name":"Institute of Emergency and Critical Care Medicine, National Yang Ming Chiao Tung University","place":["Taipei, Taiwan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5845-2235","authenticated-orcid":true,"given":"Liang","family":"Chen","sequence":"additional","affiliation":[{"name":"Division of Clinical and Translational Therapeutics, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo","place":["Buffalo, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_5_2_2","doi-asserted-by":"publisher","DOI":"10.4161\/viru.22718"},{"key":"e_1_3_5_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00093-17"},{"key":"e_1_3_5_4_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1572-0241.2005.40310.x"},{"key":"e_1_3_5_5_2","doi-asserted-by":"publisher","DOI":"10.1111\/joim.13007"},{"key":"e_1_3_5_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/iai.00508-24"},{"key":"e_1_3_5_7_2","doi-asserted-by":"publisher","DOI":"10.2147\/IJGM.S214305"},{"key":"e_1_3_5_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.micinf.2011.08.015"},{"key":"e_1_3_5_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(17)30489-9"},{"key":"e_1_3_5_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ebiom.2017.04.032"},{"key":"e_1_3_5_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jgar.2021.02.020"},{"key":"e_1_3_5_12_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2020.581543"},{"key":"e_1_3_5_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/s42003-021-02148-4"},{"key":"e_1_3_5_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.drup.2024.101137"},{"key":"e_1_3_5_15_2","doi-asserted-by":"publisher","DOI":"10.1186\/s13073-018-0587-5"},{"key":"e_1_3_5_16_2","doi-asserted-by":"publisher","DOI":"10.1007\/s10096-010-0915-1"},{"key":"e_1_3_5_17_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00089-19"},{"key":"e_1_3_5_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01750-20"},{"key":"e_1_3_5_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.00800-23"},{"key":"e_1_3_5_20_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-025-61047-y"},{"key":"e_1_3_5_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2020.01.006"},{"key":"e_1_3_5_22_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.185.3.788-800.2003"},{"key":"e_1_3_5_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.03081-22"},{"key":"e_1_3_5_24_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2021.642484"},{"key":"e_1_3_5_25_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00430-15"},{"key":"e_1_3_5_26_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00776-18"},{"key":"e_1_3_5_27_2","doi-asserted-by":"publisher","DOI":"10.1128\/iai.00559-22"},{"key":"e_1_3_5_28_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01395-10"},{"key":"e_1_3_5_29_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00843-20"},{"key":"e_1_3_5_30_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1013499"},{"key":"e_1_3_5_31_2","doi-asserted-by":"publisher","DOI":"10.1186\/s13073-025-01461-5"},{"key":"e_1_3_5_32_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00001-19"},{"key":"e_1_3_5_33_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-025-68184-4"},{"key":"e_1_3_5_34_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-micro-011420-075607"},{"key":"e_1_3_5_35_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1106733109"},{"key":"e_1_3_5_36_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.111551898"},{"key":"e_1_3_5_37_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-019-0315-1"},{"key":"e_1_3_5_38_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.lanmic.2026.101393"},{"key":"e_1_3_5_39_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.micinf.2013.10.019"},{"key":"e_1_3_5_40_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1005595"},{"key":"e_1_3_5_41_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btu153"},{"key":"e_1_3_5_42_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-24448-3"},{"key":"e_1_3_5_43_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkw1004"},{"key":"e_1_3_5_44_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.02412-14"},{"key":"e_1_3_5_45_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkv1239"},{"key":"e_1_3_5_46_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00315-09"},{"key":"e_1_3_5_47_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btr039"},{"key":"e_1_3_5_48_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0108424"},{"key":"e_1_3_5_49_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00731-18"},{"key":"e_1_3_5_50_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01950-15"},{"key":"e_1_3_5_51_2","volume-title":"Metagenomics and its applications in agriculture, biomedicine and environmental studies","author":"Salamov VSA","year":"2011","unstructured":"Salamov VSA, Solovyevand A. 2011. Automatic annotation of microbial genomes and metagenomic sequences. In Metagenomics and its applications in agriculture, biomedicine and environmental studies. Vol. 10. Nova Science Publishers."},{"key":"e_1_3_5_52_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-021-03819-2"},{"key":"e_1_3_5_53_2","first-page":"82","article-title":"Pymol: An open-source molecular graphics tool","volume":"40","author":"DeLano WL","year":"2002","unstructured":"DeLano WL. 2002. Pymol: An open-source molecular graphics tool. CCP4 Newsl protein crystallogr 40:82\u201392.","journal-title":"CCP4 Newsl protein crystallogr"},{"key":"e_1_3_5_54_2","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.1994.1017"},{"key":"e_1_3_5_55_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2021.759208"},{"key":"e_1_3_5_56_2","doi-asserted-by":"publisher","DOI":"10.1128\/jmbe.v12i1.249"},{"key":"e_1_3_5_57_2","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.02867-23"},{"key":"e_1_3_5_58_2","doi-asserted-by":"publisher","DOI":"10.1128\/mSphere.00850-19"},{"key":"e_1_3_5_59_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/bts611"},{"key":"e_1_3_5_60_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkz114"},{"key":"e_1_3_5_61_2","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-014-0550-8"},{"key":"e_1_3_5_62_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/bty895"},{"key":"e_1_3_5_63_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkab447"},{"key":"e_1_3_5_64_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.xinn.2021.100141"},{"key":"e_1_3_5_65_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkz333"}],"container-title":["Microbiology Spectrum"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.04174-25","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.04174-25","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T01:02:03Z","timestamp":1788224523000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/spectrum.04174-25"}},"subtitle":[],"editor":[{"given":"Kai","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,7,7]]},"references-count":64,"journal-issue":{"issue":"7","published-print":{"date-parts":[[2026,7,7]]}},"alternative-id":["10.1128\/spectrum.04174-25"],"URL":"https:\/\/doi.org\/10.1128\/spectrum.04174-25","relation":{},"ISSN":["2165-0497"],"issn-type":[{"value":"2165-0497","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,7]]},"assertion":[{"value":"2025-12-23","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-05-10","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-06-09","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e04174-25"}}