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In infections with\n            <jats:named-content content-type=\"genus-species\">Mycobacterium tuberculosis<\/jats:named-content>\n            , such persisters underlie the need for long-term antibiotic therapy and contribute to treatment failure in tuberculosis cases. Here, we demonstrate the value of\n            <jats:underline>d<\/jats:underline>\n            ual-\n            <jats:underline>r<\/jats:underline>\n            eporter\n            <jats:underline>m<\/jats:underline>\n            ycobacteriophages (\u03a6\n            <jats:sup>2<\/jats:sup>\n            DRMs) for characterizing\n            <jats:named-content content-type=\"genus-species\">M. tuberculosis<\/jats:named-content>\n            persisters. The addition of isoniazid (INH) to exponentially growing\n            <jats:named-content content-type=\"genus-species\">M. tuberculosis<\/jats:named-content>\n            cells consistently resulted in a 2- to 3-log decrease in CFU within 4\u00a0days, and the remaining \u22641% of cells, which survived despite being INH sensitive, were INH-tolerant persisters with a distinct transcriptional profile. We fused the promoters of several genes upregulated in persisters to the red fluorescent protein tdTomato gene in \u03a6\n            <jats:sup>2<\/jats:sup>\n            GFP10, a mycobacteriophage constitutively expressing green fluorescent protein (GFP), thus generating \u03a6\n            <jats:sup>2<\/jats:sup>\n            DRMs. A population enriched in INH persisters exhibited strong red fluorescence, by microscopy and flow cytometry, using a \u03a6\n            <jats:sup>2<\/jats:sup>\n            DRM with tdTomato controlled from the\n            <jats:italic>dnaK<\/jats:italic>\n            promoter. Interestingly, we demonstrated that, prior to INH exposure, a population primed for persistence existed in\n            <jats:named-content content-type=\"genus-species\">M. tuberculosis<\/jats:named-content>\n            cells from both cultures and human sputa and that this population was highly enriched following INH exposure. We conclude that \u03a6\n            <jats:sup>2<\/jats:sup>\n            DRMs provide a new tool to identify and quantitate\n            <jats:named-content content-type=\"genus-species\">M. tuberculosis<\/jats:named-content>\n            persister cells.\n          <\/jats:p>\n          <jats:p>\n            <jats:bold>IMPORTANCE<\/jats:bold>\n            Tuberculosis (TB) is again the leading cause of death from a single infectious disease, having surpassed HIV. The recalcitrance of the TB pandemic is largely due to the ability of the pathogen\n            <jats:named-content content-type=\"genus-species\">Mycobacterium tuberculosis<\/jats:named-content>\n            to enter a persistent state in which it is less susceptible to antibiotics and immune effectors, necessitating lengthy treatment. It has been difficult to study persister cells, as we have lacked tools to isolate these rare cells. In this article, we describe the development of dual-reporter mycobacteriophages that encode a green fluorescent marker of viability and in which the promoters of genes we have identified as induced in the persister state are fused to a gene encoding a red fluorescent protein. We show that these tools can identify heterogeneity in a cell population that correlates with propensity to survive antibiotic treatment and that the proportions of these subpopulations change in\n            <jats:named-content content-type=\"genus-species\">M. tuberculosis<\/jats:named-content>\n            cells within human sputum during the course of treatment.\n          <\/jats:p>","DOI":"10.1128\/mbio.01023-16","type":"journal-article","created":{"date-parts":[[2016,10,26]],"date-time":"2016-10-26T02:08:19Z","timestamp":1477447699000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":84,"title":["Dual-Reporter Mycobacteriophages (\u03a6\n            <sup>2<\/sup>\n            DRMs) Reveal Preexisting Mycobacterium tuberculosis Persistent Cells in Human Sputum"],"prefix":"10.1128","volume":"7","author":[{"given":"Paras","family":"Jain","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, Albert Einstein College of Medicine, New York,New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brian C.","family":"Weinrick","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Albert Einstein College of Medicine, New York,New York, USA"},{"name":"Howard Hughes Medical Institute, Albert Einstein College of Medicine, New York, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eric J.","family":"Kalivoda","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Albert Einstein College of Medicine, New York,New York, USA"},{"name":"Howard Hughes Medical Institute, Albert Einstein College of Medicine, New York, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Albert Einstein College of Medicine, New York,New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vanisha","family":"Munsamy","sequence":"additional","affiliation":[{"name":"KwaZulu-Natal Research Institute for TB\/HIV (K-RITH), Durban, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Catherine","family":"Vilcheze","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Albert Einstein College of Medicine, New York,New York, USA"},{"name":"Howard Hughes Medical Institute, Albert Einstein College of Medicine, New York, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Torin R.","family":"Weisbrod","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Albert Einstein College of Medicine, New York,New York, USA"},{"name":"Howard Hughes Medical Institute, Albert Einstein College of Medicine, New York, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michelle H.","family":"Larsen","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Albert Einstein College of Medicine, New York,New York, USA"},{"name":"Department of Medicine, Albert Einstein College of Medicine, New York, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Max R.","family":"O\u2019Donnell","sequence":"additional","affiliation":[{"name":"Division of Pulmonary, Columbia University Medical Center, New York, New York, USA"},{"name":"Department of Epidemiology Allergy and Critical Care Medicine, Columbia University Medical Center, New York, New York, USA"},{"name":"Centre for AIDS Programme of Research in South Africa (CAPRISA), Durban, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander","family":"Pym","sequence":"additional","affiliation":[{"name":"KwaZulu-Natal Research Institute for TB\/HIV 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