{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,18]],"date-time":"2026-07-18T06:49:26Z","timestamp":1784357366613,"version":"3.55.0"},"reference-count":52,"publisher":"Public Library of Science (PLoS)","issue":"2","license":[{"start":{"date-parts":[[2018,2,13]],"date-time":"2018-02-13T00:00:00Z","timestamp":1518480000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001824","name":"Grantov\u00e1 Agentura \u010cesk\u00e9 Republiky","doi-asserted-by":"publisher","award":["17-10656S"],"award-info":[{"award-number":["17-10656S"]}],"id":[{"id":"10.13039\/501100001824","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001824","name":"Grantov\u00e1 Agentura \u010cesk\u00e9 Republiky","doi-asserted-by":"publisher","award":["17-10656S"],"award-info":[{"award-number":["17-10656S"]}],"id":[{"id":"10.13039\/501100001824","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001824","name":"Grantov\u00e1 Agentura \u010cesk\u00e9 Republiky","doi-asserted-by":"publisher","award":["16-18699S"],"award-info":[{"award-number":["16-18699S"]}],"id":[{"id":"10.13039\/501100001824","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001824","name":"Grantov\u00e1 Agentura \u010cesk\u00e9 Republiky","doi-asserted-by":"publisher","award":["16-18699S"],"award-info":[{"award-number":["16-18699S"]}],"id":[{"id":"10.13039\/501100001824","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000921","name":"European Cooperation in Science and Technology ()","doi-asserted-by":"crossref","award":["CM1307"],"award-info":[{"award-number":["CM1307"]}],"id":[{"id":"10.13039\/501100000921","id-type":"DOI","asserted-by":"crossref"}]},{"name":"European Research Council ()","award":["CZ LL1601"],"award-info":[{"award-number":["CZ LL1601"]}]},{"name":"Ministerstvo \u0160kolstv\u00ed, Ml\u00e1de\u017ee a T\u011blov\u00fdchovy (CZ)","award":["LO1208 \u201cTEWEP\u201d"],"award-info":[{"award-number":["LO1208 \u201cTEWEP\u201d"]}]},{"name":"Charles University Research Center","award":["UNCE project 204072"],"award-info":[{"award-number":["UNCE project 204072"]}]},{"DOI":"10.13039\/501100006704","name":"University of Ostrava","doi-asserted-by":"crossref","award":["SGS\/PrF\/2018"],"award-info":[{"award-number":["SGS\/PrF\/2018"]}],"id":[{"id":"10.13039\/501100006704","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"DOI":"10.1371\/journal.pone.0192723","type":"journal-article","created":{"date-parts":[[2018,2,13]],"date-time":"2018-02-13T18:40:55Z","timestamp":1518547255000},"page":"e0192723","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":33,"title":["CRISPR\/Cas9 in Leishmania mexicana: A case study of LmxBTN1"],"prefix":"10.1371","volume":"13","author":[{"given":"Aygul","family":"Ishemgulova","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jana","family":"Hlav\u00e1\u010dov\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7701-5778","authenticated-orcid":true,"given":"Karolina","family":"Majerov\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anzhelika","family":"Butenko","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julius","family":"Luke\u0161","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jan","family":"Vot\u00fdpka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Petr","family":"Volf","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4765-3263","authenticated-orcid":true,"given":"Vyacheslav","family":"Yurchenko","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2018,2,13]]},"reference":[{"key":"ref1","doi-asserted-by":"crossref","first-page":"e0004349","DOI":"10.1371\/journal.pntd.0004349","article-title":"A historical overview of the classification, evolution, and dispersion of <italic>Leishmania<\/italic> parasites and sandflies","volume":"10","author":"M Akhoundi","year":"2016","journal-title":"PLOS Negl Trop Dis"},{"key":"ref2","doi-asserted-by":"crossref","first-page":"e35671","DOI":"10.1371\/journal.pone.0035671","article-title":"Leishmaniasis worldwide and global estimates of its incidence","volume":"7","author":"J Alvar","year":"2012","journal-title":"PLOS One"},{"key":"ref3","unstructured":"WHO (2016) Leishmaniasis: stuation and trends. Global health observatory (GHO) data."},{"key":"ref4","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1111\/j.1365-2915.2012.01034.x","article-title":"Phlebotomine sandflies and the spreading of leishmaniases and other diseases of public health concern","volume":"27","author":"M Maroli","year":"2013","journal-title":"Med Vet Entomol"},{"key":"ref5","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1186\/1756-3305-7-222","article-title":"Speculations on biting midges and other bloodsucking arthropods as alternative vectors of <italic>Leishmania<\/italic>","volume":"7","author":"V Seblov\u00e1","year":"2014","journal-title":"Parasit Vectors"},{"key":"ref6","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1186\/1756-3305-5-276","article-title":"<italic>Leishmania<\/italic> development in sand flies: parasite-vector interactions overview","volume":"5","author":"A Dost\u00e1lov\u00e1","year":"2012","journal-title":"Parasit Vectors"},{"key":"ref7","doi-asserted-by":"crossref","first-page":"601","DOI":"10.2174\/1566524043360285","article-title":"New insights into the developmental biology and transmission mechanisms of <italic>Leishmania<\/italic>","volume":"4","author":"PA Bates","year":"2004","journal-title":"Curr Mol Med"},{"key":"ref8","doi-asserted-by":"crossref","first-page":"e1005186","DOI":"10.1371\/journal.ppat.1005186","article-title":"Comparative life cycle transcriptomics revises <italic>Leishmania mexicana<\/italic> genome annotation and links a chromosome duplication with parasitism of vertebrates","volume":"11","author":"M Fiebig","year":"2015","journal-title":"PLOS Pathog"},{"key":"ref9","doi-asserted-by":"crossref","first-page":"23704","DOI":"10.1038\/srep23704","article-title":"Genome of <italic>Leptomonas pyrrhocoris<\/italic>: a high-quality reference for monoxenous trypanosomatids and new insights into evolution of <italic>Leishmania<\/italic>","volume":"6","author":"P Flegontov","year":"2016","journal-title":"Sci Rep"},{"key":"ref10","doi-asserted-by":"crossref","first-page":"e0005782","DOI":"10.1371\/journal.pntd.0005782","article-title":"A putative ATP\/GTP binding protein affects <italic>Leishmania mexicana<\/italic> growth in insect vectors and vertebrate hosts","volume":"11","author":"A Ishemgulova","year":"2017","journal-title":"PLOS Negl Trop Dis"},{"key":"ref11","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.pt.2014.12.012","article-title":"The past, present, and future of <italic>Leishmania<\/italic> genomics and transcriptomics","volume":"31","author":"C Cantacessi","year":"2015","journal-title":"Trends Parasitol"},{"key":"ref12","doi-asserted-by":"crossref","first-page":"e0154101","DOI":"10.1371\/journal.pone.0154101","article-title":"Genomic and molecular characterization of miltefosine resistance in <italic>Leishmania<\/italic> infantum strains with either natural or acquired resistance through experimental selection of intracellular amastigotes","volume":"11","author":"A Mondelaers","year":"2016","journal-title":"PLOS One"},{"key":"ref13","doi-asserted-by":"crossref","first-page":"15458","DOI":"10.1073\/pnas.2136651100","article-title":"A role in vacuolar arginine transport for yeast Btn1p and for human CLN3, the protein defective in Batten disease","volume":"100","author":"Y Kim","year":"2003","journal-title":"Proc Natl Acad Sci U S A"},{"key":"ref14","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1038\/8861","article-title":"Action of <italic>BTN1<\/italic>, the yeast orthologue of the gene mutated in Batten disease","volume":"22","author":"DA Pearce","year":"1999","journal-title":"Nat Genet"},{"key":"ref15","doi-asserted-by":"crossref","first-page":"10273","DOI":"10.1074\/jbc.M510625200","article-title":"<italic>Saccharomyces cerevisiae<\/italic> lacking <italic>Btn1p<\/italic> modulate vacuolar ATPase activity to regulate pH imbalance in the vacuole","volume":"281","author":"S Padilla-Lopez","year":"2006","journal-title":"J Biol Chem"},{"key":"ref16","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1242\/dmm.008490","article-title":"<italic>BTN1<\/italic>, the <italic>Saccharomyces cerevisiae<\/italic> homolog to the human Batten disease gene, is involved in phospholipid distribution","volume":"5","author":"S Padilla-Lopez","year":"2012","journal-title":"Dis Model Mech"},{"key":"ref17","doi-asserted-by":"crossref","first-page":"1313","DOI":"10.1091\/mbc.E03-02-0120","article-title":"Two motifs target Batten disease protein CLN3 to lysosomes in transfected nonneuronal and neuronal cells","volume":"15","author":"A Kyttala","year":"2004","journal-title":"Mol Biol Cell"},{"key":"ref18","doi-asserted-by":"crossref","first-page":"573","DOI":"10.1002\/jnr.20367","article-title":"CLN3, the protein associated with batten disease: structure, function and localization","volume":"79","author":"SN Phillips","year":"2005","journal-title":"J Neurosci Res"},{"key":"ref19","doi-asserted-by":"crossref","first-page":"D457","DOI":"10.1093\/nar\/gkp851","article-title":"TriTrypDB: a functional genomic resource for the Trypanosomatidae","volume":"38","author":"M Aslett","year":"2010","journal-title":"Nucleic Acids Res"},{"key":"ref20","doi-asserted-by":"crossref","first-page":"1792","DOI":"10.1093\/nar\/gkh340","article-title":"MUSCLE: multiple sequence alignment with high accuracy and high throughput","volume":"32","author":"RC Edgar","year":"2004","journal-title":"Nucleic Acids Res"},{"key":"ref21","doi-asserted-by":"crossref","first-page":"564","DOI":"10.1080\/10635150701472164","article-title":"Improvement of phylogenies after removing divergent and ambiguously aligned blocks from protein sequence alignments","volume":"56","author":"G Talavera","year":"2007","journal-title":"Syst Biol"},{"key":"ref22","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1093\/molbev\/msu300","article-title":"IQ-TREE: a fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies","volume":"32","author":"LT Nguyen","year":"2015","journal-title":"Mol Biol Evol"},{"key":"ref23","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1093\/sysbio\/sys029","article-title":"MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space","volume":"61","author":"F Ronquist","year":"2012","journal-title":"Syst Biol"},{"key":"ref24","doi-asserted-by":"crossref","first-page":"e1005127","DOI":"10.1371\/journal.ppat.1005127","article-title":"<italic>Leptomonas seymouri<\/italic>: adaptations to the dixenous life cycle analyzed by genome sequencing, transcriptome profiling and co-infection with <italic>Leishmania donovani<\/italic>","volume":"11","author":"N Kraeva","year":"2015","journal-title":"PLOS Pathog"},{"issue":"Pt 1","key":"ref25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1017\/S0031182000078458","article-title":"Complete developmental cycle of <italic>Leishmania mexicana<\/italic> in axenic culture","volume":"108","author":"PA Bates","year":"1994","journal-title":"Parasitology"},{"key":"ref26","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1186\/1471-2164-9-255","article-title":"Genome-wide gene expression profiling analysis of <italic>Leishmania major<\/italic> and <italic>Leishmania infantum<\/italic> developmental stages reveals substantial differences between the two species","volume":"9","author":"A Rochette","year":"2008","journal-title":"BMC Genomics"},{"key":"ref27","doi-asserted-by":"crossref","first-page":"1765","DOI":"10.1111\/j.1462-5822.2010.01507.x","article-title":"The stage-regulated HASPB and SHERP proteins are essential for differentiation of the protozoan parasite <italic>Leishmania major<\/italic> in its sand fly vector, <italic>Phlebotomus papatasi<\/italic>","volume":"12","author":"J S\u00e1dlov\u00e1","year":"2010","journal-title":"Cell Microbiol"},{"key":"ref28","doi-asserted-by":"crossref","first-page":"016","DOI":"10.14411\/fp.2016.016","article-title":"T7 polymerase-driven transcription is downregulated in metacyclic promastigotes and amastigotes of <italic>Leishmania mexicana<\/italic>","volume":"63","author":"A Ishemgulova","year":"2016","journal-title":"Folia Parasitol"},{"key":"ref29","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1016\/j.febslet.2014.01.034","article-title":"A small portion of plastid transcripts is polyadenylated in the flagellate <italic>Euglena gracilis<\/italic>","volume":"588","author":"K Z\u00e1honov\u00e1","year":"2014","journal-title":"FEBS Lett"},{"key":"ref30","doi-asserted-by":"crossref","first-page":"1940","DOI":"10.3389\/fmicb.2017.01940","article-title":"Genome of <italic>Ca<\/italic>. Pandoraea novymonadis, an endosymbiotic bacterium of the trypanosomatid <italic>Novymonas esmeralda<\/italic>","volume":"8","author":"AY Kostygov","year":"2017","journal-title":"Front Microbiol"},{"key":"ref31","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1016\/j.molbiopara.2013.05.002","article-title":"Identification of essential and non-essential protein kinases by a fusion PCR method for efficient production of transgenic <italic>Trypanosoma brucei<\/italic>","volume":"190","author":"C Merritt","year":"2013","journal-title":"Mol Biochem Parasitol"},{"key":"ref32","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.pep.2005.03.004","article-title":"Development of an inducible protein expression system based on the protozoan host <italic>Leishmania tarentolae<\/italic>","volume":"42","author":"S Kushnir","year":"2005","journal-title":"Protein Expres Purif"},{"key":"ref33","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.molbiopara.2014.11.002","article-title":"Tetracycline-inducible gene expression system in <italic>Leishmania mexicana<\/italic>","volume":"198","author":"N Kraeva","year":"2014","journal-title":"Mol Biochem Parasitol"},{"key":"ref34","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1016\/S0022-2836(75)80083-0","article-title":"Detection of specific sequences among DNA fragments separated by gel electrophoresis","volume":"98","author":"EM Southern","year":"1975","journal-title":"J Mol Biol"},{"key":"ref35","doi-asserted-by":"crossref","first-page":"819","DOI":"10.1126\/science.1231143","article-title":"Multiplex genome engineering using CRISPR\/Cas systems","volume":"339","author":"L Cong","year":"2013","journal-title":"Science"},{"key":"ref36","doi-asserted-by":"crossref","first-page":"6736","DOI":"10.1128\/MCB.14.10.6736","article-title":"Upstream tRNA genes are essential for expression of small nuclear and cytoplasmic RNA genes in trypanosomes","volume":"14","author":"V Nakaar","year":"1994","journal-title":"Mol Cell Biol"},{"key":"ref37","first-page":"e000033","article-title":"EuPaGDT: a web tool tailored to design CRISPR guide RNAs for eukaryotic pathogens","volume":"1","author":"D Peng","year":"2015","journal-title":"Microb Genom"},{"key":"ref38","doi-asserted-by":"crossref","first-page":"1405","DOI":"10.1111\/cmi.12456","article-title":"First efficient CRISPR-Cas9-mediated genome editing in <italic>Leishmania<\/italic> parasites","volume":"17","author":"L Sollelis","year":"2015","journal-title":"Cell Microbiol"},{"key":"ref39","first-page":"S1","article-title":"Establishment and maintenance of sand fly colonies","volume":"1","author":"P Volf","year":"2011","journal-title":"J Vector Ecol 36 Suppl"},{"key":"ref40","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1093\/jmedent\/45.1.133","article-title":"<italic>Leishmania<\/italic> in sand flies: comparison of quantitative polymerase chain reaction with other techniques to determine the intensity of infection","volume":"45","author":"J My\u0161kov\u00e1","year":"2008","journal-title":"J Med Entomol"},{"key":"ref41","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1017\/S0031182002001439","article-title":"The role of promastigote secretory gel in the origin and transmission of the infective stage of <italic>Leishmania mexicana<\/italic> by the sandfly <italic>Lutzomyia longipalpis<\/italic>","volume":"124","author":"ME Rogers","year":"2002","journal-title":"Parasitology"},{"key":"ref42","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1186\/s13071-015-0765-x","article-title":"Xenodiagnosis of <italic>Leishmania donovani<\/italic> in BALB\/c mice using <italic>Phlebotomus orientalis<\/italic>: a new laboratory model","volume":"8","author":"J S\u00e1dlov\u00e1","year":"2015","journal-title":"Parasit Vectors"},{"key":"ref43","doi-asserted-by":"crossref","first-page":"537","DOI":"10.1111\/j.1550-7408.2012.00636.x","article-title":"New species of insect trypanosomatids from Costa Rica and the proposal for a new subfamily within the Trypanosomatidae","volume":"59","author":"M Jirk\u016f","year":"2012","journal-title":"J Eukaryot Microbiol"},{"key":"ref44","doi-asserted-by":"crossref","first-page":"1131","DOI":"10.1093\/nar\/gkr834","article-title":"Genome sequencing of the lizard parasite <italic>Leishmania tarentolae<\/italic> reveals loss of genes associated to the intracellular stage of human pathogenic species","volume":"40","author":"F Raymond","year":"2011","journal-title":"Nucleic Acids Res"},{"key":"ref45","doi-asserted-by":"crossref","first-page":"2350","DOI":"10.12688\/f1000research.9218.1","article-title":"Plasticity of the <italic>Leishmania<\/italic> genome leading to gene copy number variations and drug resistance","volume":"5","author":"MN Laffitte","year":"2016","journal-title":"F1000Res"},{"key":"ref46","doi-asserted-by":"crossref","first-page":"e02097","DOI":"10.1128\/mBio.02097-14","article-title":"CRISPR-Cas9-mediated single-gene and gene family disruption in <italic>Trypanosoma cruzi<\/italic>","volume":"6","author":"D Peng","year":"2014","journal-title":"mBio"},{"key":"ref47","doi-asserted-by":"crossref","first-page":"170095","DOI":"10.1098\/rsos.170095","article-title":"A CRISPR Cas9 high-throughput genome editing toolkit for kinetoplastids","volume":"4","author":"T Beneke","year":"2017","journal-title":"R Soc Open Sci"},{"key":"ref48","doi-asserted-by":"crossref","first-page":"e00861","DOI":"10.1128\/mBio.00861-15","article-title":"CRISPR-Cas9-mediated genome editing in <italic>Leishmania donovani<\/italic>","volume":"6","author":"WW Zhang","year":"2015","journal-title":"mBio"},{"key":"ref49","doi-asserted-by":"crossref","first-page":"e00340","DOI":"10.1128\/mSphere.00340-16","article-title":"Optimized CRISPR-Cas9 genome editing for <italic>Leishmania<\/italic> and its use to target a multigene family, induce chromosomal translocation, and study DNA break repair mechanisms","volume":"2","author":"WW Zhang","year":"2017","journal-title":"mSphere"},{"key":"ref50","doi-asserted-by":"crossref","first-page":"2619","DOI":"10.1093\/emboj\/17.9.2619","article-title":"A mitogen-activated protein (MAP) kinase homologue of <italic>Leishmania mexicana<\/italic> is essential for parasite survival in the infected host","volume":"17","author":"M Wiese","year":"1998","journal-title":"EMBO J"},{"key":"ref51","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/S0166-6851(97)00078-9","article-title":"Structure of the <italic>Trypanosoma brucei<\/italic> U6 snRNA gene promoter","volume":"88","author":"V Nakaar","year":"1997","journal-title":"Mol Biochem Parasitol"},{"key":"ref52","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1080\/00034983.1999.11813463","article-title":"<italic>Leishmania major<\/italic>: effect of repeated passages through sandfly vectors or murine hosts","volume":"93","author":"J S\u00e1dlov\u00e1","year":"1999","journal-title":"Ann Trop Med Parasitol"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/dx.plos.org\/10.1371\/journal.pone.0192723","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,5,19]],"date-time":"2020-05-19T00:47:55Z","timestamp":1589849275000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0192723"}},"subtitle":[],"editor":[{"given":"Andrew Paul","family":"Jackson","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2018,2,13]]},"references-count":52,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2018,2,13]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0192723","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,2,13]]}}}