{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,21]],"date-time":"2026-04-21T14:27:43Z","timestamp":1776781663929,"version":"3.51.2"},"reference-count":41,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2022,11,18]],"date-time":"2022-11-18T00:00:00Z","timestamp":1668729600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000024","name":"Canadian Institutes of Health Research","doi-asserted-by":"publisher","award":["125996"],"award-info":[{"award-number":["125996"]}],"id":[{"id":"10.13039\/501100000024","id-type":"DOI","asserted-by":"publisher"}]},{"name":"R. Howard Webster Foundation","award":["125996"],"award-info":[{"award-number":["125996"]}]},{"name":"Foundation of the McGill University Health Center","award":["125996"],"award-info":[{"award-number":["125996"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["TropicalMed"],"abstract":"<jats:p>The protozoan parasite Leishmania donovani is a causative agent of the neglected tropical disease known as visceral leishmaniasis, which can be lethal when untreated. Studying Leishmania viru-lence factors is crucial in determining how the parasite causes disease and identifying new targets for treatment. One potential virulence factor is L. donovani\u2019s abundantly secreted protein: secreted acid phosphatase (SAcP). Whole-genome analysis revealed that the sacp gene was present in three copies in wild type L. donovani. Using CRISPR-Cas9 gene editing; we generated a sacp gene knockout termed Ld\u0394SAcP, which demonstrated a loss of both the SAcP protein and an associated reduction in secreted acid phosphatase activity. Genome sequencing confirmed the precise dele-tion of the sacp gene in Ld\u0394SAcP and identified several changes in the genome. Ld\u0394SAcP demonstrated no significant changes in promastigote proliferation or its ability to infect and survive in macrophages compared to the wildtype strain. Ld\u0394SAcP also demonstrated no change in murine liver infection; however, survival was impaired in the spleen. Taken together these results show that SAcP is not necessary for the survival of promastigotes in culture but may support long-term survival in the spleen. These observations also show that the use of CRISPR gene editing and WGS together are effective to investigate the function and phenotype of complex potential drug targets such as multicopy genes.<\/jats:p>","DOI":"10.3390\/tropicalmed7110384","type":"journal-article","created":{"date-parts":[[2022,11,21]],"date-time":"2022-11-21T04:13:36Z","timestamp":1669004016000},"page":"384","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Investigating the Leishmania donovani\u00a0sacp Gene and Its Role in Macrophage Infection and Survival in Mice"],"prefix":"10.3390","volume":"7","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9379-9509","authenticated-orcid":false,"given":"Kayla","family":"Paulini","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, McGill University, Montreal, QC H3A 2B4, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5550-712X","authenticated-orcid":false,"given":"Patrick","family":"Lypaczewski","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, McGill University, Montreal, QC H3A 2B4, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1996-876X","authenticated-orcid":false,"given":"Wen-Wei","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, McGill University, Montreal, QC H3A 2B4, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0874-7251","authenticated-orcid":false,"given":"Dilhan J.","family":"Perera","sequence":"additional","affiliation":[{"name":"Division of Experimental Medicine, McGill University, Montreal, QC H4A 3J1, Canada"},{"name":"Infectious Diseases and Immunity in Global Health Program, Research Institute of the McGill University Health Centre, Montreal, QC H4A 3J1, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Momar","family":"Ndao","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, McGill University, Montreal, QC H3A 2B4, Canada"},{"name":"Division of Experimental Medicine, McGill University, Montreal, QC H4A 3J1, Canada"},{"name":"Infectious Diseases and Immunity in Global Health Program, Research Institute of the McGill University Health Centre, Montreal, QC H4A 3J1, Canada"},{"name":"National Reference Centre for Parasitology, Research Institute of the McGill University Health Centre, Montreal, QC H4A 3J1, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Greg","family":"Matlashewski","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, McGill University, Montreal, QC H3A 2B4, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,11,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"951","DOI":"10.1016\/S0140-6736(18)31204-2","article-title":"Leishmaniasis","volume":"392","author":"Burza","year":"2018","journal-title":"Lancet"},{"key":"ref_2","unstructured":"CDC (2022, January 26). 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