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Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2025,6,17]]},"abstract":"<jats:p>\n                    <jats:italic toggle=\"yes\">Mycobacterium leprae<\/jats:italic>\n                    , the causative agent of leprosy, has never been cultured in vitro, posing significant challenges for genetic manipulation and drug discovery. Current antileprosy drug screening methods relying on microscopic count, radiorespirometry, and qPCR are time consuming and require the use of radioactive elements. We demonstrate mycobacteriophage-mediated introduction of foreign DNA using the broad-host range mycobacteriophage TM4 and the application of the luciferase reporter mycobacteriophage (LRM) for drug screening. Mycobacteriophage infection of\n                    <jats:italic toggle=\"yes\">M. leprae<\/jats:italic>\n                    was shown using TM4 expressing the highly sensitive BRET-nanoluciferase-based reporter, GeNL (TM4\n                    <jats:italic toggle=\"yes\">::GeNL<\/jats:italic>\n                    ), which enables luminescence measurement for over 72 h. When\n                    <jats:italic toggle=\"yes\">M. leprae<\/jats:italic>\n                    was exposed to rifampicin, dapsone, and Q203 for 24 and 48 h, followed by TM4\n                    <jats:italic toggle=\"yes\">::GeNL<\/jats:italic>\n                    infection, the luminescence output decreased in a dose-dependent manner, establishing an in vitro two-day screening assay for drugs. We have also electroporated\n                    <jats:italic toggle=\"yes\">M. leprae<\/jats:italic>\n                    with a\n                    <jats:italic toggle=\"yes\">ColE1<\/jats:italic>\n                    -integration proficient plasmid expressing GeNL and shown that the transformed leprosy bacilli could be propagated in mice footpads and detected using an in vivo imaging system (IVIS). These findings introduce powerful genetic tools for\n                    <jats:italic toggle=\"yes\">M. leprae<\/jats:italic>\n                    research enabling in vivo tracking and in vitro viability testing.\n                  <\/jats:p>","DOI":"10.1073\/pnas.2508271122","type":"journal-article","created":{"date-parts":[[2025,6,11]],"date-time":"2025-06-11T11:13:57Z","timestamp":1749640437000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":2,"title":["Mycobacteriophage-mediated gene transfer enables in vitro drug screening and in vivo tracking of\n                    <i>Mycobacterium leprae<\/i>"],"prefix":"10.1073","volume":"122","author":[{"given":"Lahari","family":"Das","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, Albert Einstein College of Medicine","place":["Bronx"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bing","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Albert Einstein College of Medicine","place":["Bronx"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Saranathan","family":"Rajagopalan","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Albert Einstein College of Medicine","place":["Bronx"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Catherine","family":"Vilcheze","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Albert Einstein College of Medicine","place":["Bronx"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claire V.","family":"Mulholland","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Albert Einstein College of Medicine","place":["Bronx"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4833-0573","authenticated-orcid":false,"given":"Michael","family":"Berney","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Albert Einstein College of Medicine","place":["Bronx"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick K.","family":"Andrews","sequence":"additional","affiliation":[{"name":"United States Department of Health and Human Services, Health Resources and Services Administration, Health Systems Bureau, National Hansen\u2019s Disease Program","place":["Baton Rouge"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashley","family":"Edwards","sequence":"additional","affiliation":[{"name":"United States Department of Health and Human Services, Health Resources and Services Administration, Health Systems Bureau, National Hansen\u2019s Disease Program","place":["Baton Rouge"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ramanuj","family":"Lahiri","sequence":"additional","affiliation":[{"name":"United States Department of Health and Human Services, Health Resources and Services Administration, Health Systems Bureau, National Hansen\u2019s Disease Program","place":["Baton Rouge"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3321-3080","authenticated-orcid":false,"suffix":"Jr.","given":"William R.","family":"Jacobs","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Albert Einstein College of Medicine","place":["Bronx"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2025,6,11]]},"reference":[{"key":"e_1_3_5_1_2","unstructured":"WHO Global leprosy (Hansen disease) update 2023: Elimination of leprosy disease is possible\u2013Time to act! 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