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Talents","award":["2021XG001"],"award-info":[{"award-number":["2021XG001"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Toxoplasma gondii, a pathogenic apicomplexan parasite, infects approximately one third of the world\u2019s population and poses a serious threat to global public health. Microneme proteins (MICs) secreted by the microneme, an apical secretory organelle of T. gondii, play important roles in the invasion, motility, and intracellular survival of T. gondii. In this study, we selected 11 genes of interest (GOIs) of T. gondii, tentative MICs predicted to be localized in micronemes, and we used the CRISPR-Cas9 system to construct epitope tagging strains and gene knockout strains to explore the localization and function of these 11 tentative MICs. Immunofluorescence assay showed that nine tentative MICs (TGME49_243930, TGME49_200270, TGME49_273320, TGME49_287040, TGME49_261710, TGME49_205680, TGME49_304490, TGME49_245485, and TGME49_224620) were localized or partially localized in the microneme, consistent with the prediction. However, TGME49_272380 and TGME49_243790 showed different localizations from the prediction, being localized in the endoplasmic reticulum and the dense granule, respectively. Further functional characterization of the 11 RH\u0394GOI strains revealed that deletion of these 11 GOIs had no significant effect on plaque formation, intracellular replication, egress, invasion ability, and virulence of T. gondii. Although these 11 GOIs are not essential genes for the growth and virulence of tachyzoites of type I RH strain, they may have potential roles in other developmental stages or other genotypes of T. gondii. Thus, further research should be performed to explore the possible role of the nine mics and the other two GOIs in other life cycle stages and other genotypes of T. gondii.<\/jats:p>","DOI":"10.3390\/ani14172543","type":"journal-article","created":{"date-parts":[[2024,9,2]],"date-time":"2024-09-02T06:51:17Z","timestamp":1725259877000},"page":"2543","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Functional Characterization of 11 Tentative Microneme Proteins in Type I RH Strain of Toxoplasma gondii Using the CRISPR-Cas9 System"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-9676-2631","authenticated-orcid":false,"given":"Zhi-Ya","family":"Ma","sequence":"first","affiliation":[{"name":"Laboratory of Parasitic Diseases, College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong 030801, China"},{"name":"State Key Laboratory for Animal Disease Control and Prevention, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiao-Jing","family":"Wu","sequence":"additional","affiliation":[{"name":"Laboratory of Parasitic Diseases, College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong 030801, China"},{"name":"State Key Laboratory for Animal Disease Control and Prevention, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chuan","family":"Li","sequence":"additional","affiliation":[{"name":"Laboratory of Parasitic Diseases, College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong 030801, China"},{"name":"State Key Laboratory for Animal Disease Control and Prevention, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jin","family":"Gao","sequence":"additional","affiliation":[{"name":"Laboratory of Parasitic Diseases, College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong 030801, China"},{"name":"State Key Laboratory for Animal Disease Control and Prevention, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-1569-3890","authenticated-orcid":false,"given":"Yong-Jie","family":"Kou","sequence":"additional","affiliation":[{"name":"Laboratory of Parasitic Diseases, College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong 030801, China"},{"name":"State Key Laboratory for Animal Disease Control and Prevention, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Meng","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2530-4628","authenticated-orcid":false,"given":"Xing-Quan","family":"Zhu","sequence":"additional","affiliation":[{"name":"Laboratory of Parasitic Diseases, College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong 030801, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiao-Nan","family":"Zheng","sequence":"additional","affiliation":[{"name":"Laboratory of Parasitic Diseases, College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong 030801, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,9,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Dubey, J.P. (2022). Toxoplasmosis of Animals and Humans, CRC Press. [3rd ed.].","DOI":"10.1201\/9781003199373"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"3095","DOI":"10.1080\/21505594.2021.2012346","article-title":"The pathogenicity and virulence of Toxoplasma gondii","volume":"12","author":"Sanchez","year":"2021","journal-title":"Virulence"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e00115-19","DOI":"10.1128\/CMR.00115-19","article-title":"Epidemiology, pathophysiology, diagnosis, and management of cerebral toxoplasmosis","volume":"34","author":"Elsheikha","year":"2021","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"959","DOI":"10.1016\/j.pt.2020.08.005","article-title":"Toxoplasma gondii: An underestimated threat?","volume":"36","author":"Milne","year":"2020","journal-title":"Trends Parasitol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"273","DOI":"10.2147\/DDDT.S60973","article-title":"Drugs in development for toxoplasmosis: Advances, challenges, and current status","volume":"11","author":"Alday","year":"2017","journal-title":"Drug Des. Devel. Ther."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e00057-17","DOI":"10.1128\/CMR.00057-17","article-title":"Treatment of toxoplasmosis: Historical perspective, animal models, and current clinical practice","volume":"31","author":"Dunay","year":"2018","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e113155","DOI":"10.15252\/embj.2022113155","article-title":"A conserved complex of microneme proteins mediates rhoptry discharge in Toxoplasma","volume":"42","author":"Valleau","year":"2023","journal-title":"EMBO J."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Lai, B.S., Juhas, M., and Zhang, Y. (2019). Toxoplasma gondii secretory proteins and their role in invasion and pathogenesis. Microbiol. Res., 227.","DOI":"10.1016\/j.micres.2019.06.003"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/S1471-4922(00)01761-X","article-title":"Mix and match modules: Structure and function of microneme proteins in apicomplexan parasites","volume":"17","author":"Tomley","year":"2001","journal-title":"Trends Parasitol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"421","DOI":"10.1046\/j.1365-2958.1999.01174.x","article-title":"Mobilization of intracellular calcium stimulates microneme discharge in Toxoplasma gondii","volume":"31","author":"Carruthers","year":"1999","journal-title":"Mol. Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/j.exppara.2017.03.001","article-title":"Research advances in interactions related to Toxoplasma gondii microneme proteins","volume":"176","author":"Liu","year":"2017","journal-title":"Exp. Parasitol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"4119","DOI":"10.1074\/jbc.M008294200","article-title":"The Toxoplasma micronemal protein MIC4 is an adhesin composed of six conserved apple domains","volume":"276","author":"Brecht","year":"2001","journal-title":"J. Biol. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Zhu, J., Wang, Y., Cao, Y., Shen, J., and Yu, L. (2021). Diverse roles of TgMIC1\/4\/6 in the Toxoplasma infection. Front. Microbiol., 12.","DOI":"10.3389\/fmicb.2021.666506"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1083\/jcb.152.3.563","article-title":"Identification and characterization of an escorter for two secretory adhesins in Toxoplasma gondii","volume":"152","author":"Reiss","year":"2001","journal-title":"J. Cell Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1084\/jem.20041672","article-title":"Synergistic role of micronemal proteins in Toxoplasma gondii virulence","volume":"201","author":"Dubremetz","year":"2005","journal-title":"J. Exp. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"4551","DOI":"10.1091\/mbc.e06-01-0064","article-title":"A cleavable propeptide influences Toxoplasma infection by facilitating the trafficking and secretion of the TgMIC2-M2AP invasion complex","volume":"17","author":"Harper","year":"2006","journal-title":"Mol. Biol. Cell"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2082","DOI":"10.1093\/emboj\/cdg217","article-title":"Rapid invasion of host cells by Toxoplasma requires secretion of the MIC2-M2AP adhesive protein complex","volume":"22","author":"Huynh","year":"2003","journal-title":"EMBO J."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1242\/jcs.115.3.563","article-title":"A family of transmembrane microneme proteins of Toxoplasma gondii contain EGF-like domains and function as escorters","volume":"115","author":"Meissner","year":"2002","journal-title":"J. Cell Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"947","DOI":"10.1242\/jcs.022350","article-title":"Microneme protein 8--a new essential invasion factor in Toxoplasma gondii","volume":"121","author":"Kessler","year":"2008","journal-title":"J. Cell Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"752","DOI":"10.1016\/j.chom.2020.09.011","article-title":"A comprehensive subcellular atlas of the Toxoplasma proteome via hyperLOPIT provides spatial context for protein functions","volume":"28","author":"Barylyuk","year":"2020","journal-title":"Cell Host Microbe"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"3165","DOI":"10.1096\/fj.201901416RR","article-title":"Novel roles of dense granule protein 12 (GRA12) in Toxoplasma gondii infection","volume":"34","author":"Wang","year":"2020","journal-title":"FASEB J."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Liang, Q.L., Nie, L.B., Li, T.T., Elsheikha, H.M., Sun, L.X., Zhang, Z.W., Zhao, D.Y., Zhu, X.Q., and Wang, J.L. (2021). Functional characterization of 17 protein serine\/threonine phosphatases in Toxoplasma gondii using CRISPR-Cas9 system. Front. Cell Dev. Biol., 9.","DOI":"10.3389\/fcell.2021.738794"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Zheng, X.N., Wang, J.L., Elsheikha, H.M., Wang, M., Zhang, Z.W., Sun, L.X., Wang, X.C., Zhu, X.Q., and Li, T.T. (2023). Functional characterization of 15 novel dense granule proteins in Toxoplasma gondii using the CRISPR-Cas9 system. Microbiol. Spectr., 11.","DOI":"10.1128\/spectrum.03078-22"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Zhang, Z.W., Li, T.T., Wang, J.L., Liang, Q.L., Zhang, H.S., Sun, L.X., and Zhu, X.Q. (2020). Functional characterization of two thioredoxin proteins of Toxoplasma gondii using the CRISPR-Cas9 system. Front. Vet. Sci., 7.","DOI":"10.3389\/fvets.2020.614759"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/j.ijpara.2023.09.001","article-title":"A newly characterized dense granule protein (GRA76) is important for the growth and virulence of Toxoplasma gondii","volume":"54","author":"Zheng","year":"2024","journal-title":"Int. J. Parasitol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Wu, X.J., Gao, J., Zheng, X.N., Elsheikha, H.M., Li, T.T., Kou, Y.J., Wang, M., and Zhu, X.Q. (2023). The splicing factor SR2 is an important virulence factor of Toxoplasma gondii. Front. Microbiol., 14.","DOI":"10.3389\/fmicb.2023.1302512"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Gao, J., Wu, X.J., Zheng, X.N., Li, T.T., Kou, Y.J., Wang, X.C., Wang, M., and Zhu, X.Q. (2023). Functional characterization of eight zinc finger motif-containing proteins in Toxoplasma gondii Type I RH strain using the CRISPR-Cas9 system. Pathogens, 12.","DOI":"10.3390\/pathogens12101232"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1007\/s00431-005-1665-4","article-title":"Treatment of infants with congenital toxoplasmosis: Tolerability and plasma concentrations of sulfadiazine and pyrimethamine","volume":"165","author":"Schmidt","year":"2006","journal-title":"Eur. J. Pediatr."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Montazeri, M., Sharif, M., Sarvi, S., Mehrzadi, S., Ahmadpour, E., and Daryani, A. (2017). A systematic review of in vitro and in vivo activities of anti-Toxoplasma drugs and compounds (2006\u20132016). Front. Microbiol., 8.","DOI":"10.3389\/fmicb.2017.00025"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1007\/s10096-018-03442-6","article-title":"A systematic review on efficiency of microneme proteins to induce protective immunity against Toxoplasma gondii","volume":"38","author":"Dodangeh","year":"2019","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"ref_31","first-page":"193","article-title":"Congenital toxoplasmosis treatment","volume":"15","author":"Serranti","year":"2011","journal-title":"Eur. Rev. Med. Pharmacol. Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"108385","DOI":"10.1016\/j.exppara.2022.108385","article-title":"Protective immunity induced with a DNA vaccine encoding B- and T-cells multi-epitope SAG1, ROP16, MIC4, GRA12, M2AP, and multi-epitope ROP8 against acute and chronic toxoplasmosis in BALB\/c mice","volume":"242","author":"Ghaffari","year":"2022","journal-title":"Exp. Parasitol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"526","DOI":"10.1007\/s12639-013-0390-6","article-title":"A comparative study between excretory\/secretory and autoclaved vaccines against RH strain of Toxoplasma gondii in murine models","volume":"39","author":"Kamel","year":"2015","journal-title":"J. Parasit. Dis."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Dubois, D.J., and Soldati-Favre, D. (2019). Biogenesis and secretion of micronemes in Toxoplasma gondii. Cell. Microbiol., 21.","DOI":"10.1111\/cmi.13018"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1702","DOI":"10.1292\/jvms.18-0344","article-title":"How does Toxoplasma gondii invade host cells?","volume":"80","author":"Kato","year":"2018","journal-title":"J. Vet. Med. Sci."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Lamarque, M., Besteiro, S., Papoin, J., Roques, M., Vulliez-Le Normand, B., Morlon-Guyot, J., Dubremetz, J.F., Fauquenoy, S., Tomavo, S., and Faber, B.W. (2011). The RON2-AMA1 interaction is a critical step in moving junction-dependent invasion by apicomplexan parasites. PloS Pathog., 7.","DOI":"10.1371\/journal.ppat.1001276"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"619","DOI":"10.1146\/annurev-micro-041320-021425","article-title":"How apicomplexa parasites secrete and build their invasion machinery","volume":"76","author":"Cova","year":"2022","journal-title":"Annu. Rev. Microbiol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1423","DOI":"10.1016\/j.cell.2016.08.019","article-title":"A genome-wide CRISPR screen in Toxoplasma identifies essential apicomplexan genes","volume":"166","author":"Sidik","year":"2016","journal-title":"Cell"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"E6956","DOI":"10.1073\/pnas.1722202115","article-title":"Human impact on the diversity and virulence of the ubiquitous zoonotic parasite Toxoplasma gondii","volume":"115","author":"Shwab","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1038\/s41579-021-00518-7","article-title":"Toxoplasma gondii infection and its implications within the central nervous system","volume":"19","author":"Matta","year":"2021","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1084\/jem.20100717","article-title":"Strain-specific activation of the NF-kappaB pathway by GRA15, a novel Toxoplasma gondii dense granule protein","volume":"208","author":"Rosowski","year":"2011","journal-title":"J. Exp. 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