{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,30]],"date-time":"2026-08-30T02:48:20Z","timestamp":1788058100298,"version":"build-2784847793"},"reference-count":23,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2024,2,23]],"date-time":"2024-02-23T00:00:00Z","timestamp":1708646400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/100015330","name":"Max Kade foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100015330","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000781","name":"European Research Council","doi-asserted-by":"publisher","award":["ERC StG 281719"],"award-info":[{"award-number":["ERC StG 281719"]}],"id":[{"id":"10.13039\/501100000781","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["SFB 1129"],"award-info":[{"award-number":["SFB 1129"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["SPP 2225"],"award-info":[{"award-number":["SPP 2225"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["SPP 2332"],"award-info":[{"award-number":["SPP 2332"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000854","name":"Human Frontier Science Program","doi-asserted-by":"publisher","award":["RGY0071\/2011"],"award-info":[{"award-number":["RGY0071\/2011"]}],"id":[{"id":"10.13039\/501100000854","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Molecular Microbiology"],"published-print":{"date-parts":[[2024,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Plasmodium sporozoites are the highly motile and invasive forms of the malaria parasite transmitted by mosquitoes. Sporozoites form within oocysts at the midgut wall of the mosquito, egress from oocysts and enter salivary glands prior to transmission. The GPI\u2010anchored major surface protein, the circumsporozoite protein (CSP) is important for <jats:italic>Plasmodium<\/jats:italic> sporozoite formation, egress, migration and invasion. To visualize CSP, we previously generated full\u2010length versions of CSP internally tagged with the green fluorescent protein, GFP. However, while these allowed for imaging of sporogony in oocysts, sporozoites failed to egress. Here, we explore different strategies to overcome this block in egress and obtain salivary gland resident sporozoites that express CSP\u2010GFP. Replacing the N\u2010terminal and repeat region with GFP did not allow sporozoite formation. Lowering expression of CSP\u2010GFP at the endogenous locus allowed sporozoite formation but did not overcome egress block. Crossing of CSP\u2010GFP expressing parasites that are blocked in egress with wild\u2010type parasites yielded a small fraction of parasites that entered salivary glands and expressed various levels of CSP\u2010GFP. Expressing CSP\u2010GFP constructs from a silent chromosome region from promoters that are active only post salivary gland invasion yielded normal numbers of fluorescent salivary gland sporozoites, albeit with low levels of fluorescence. We also show that lowering CSP expression by 50% allowed egress from oocysts but not salivary gland entry. In conclusion, <jats:italic>Plasmodium berghei<\/jats:italic> parasites with normal CSP expression tolerate a certain level of CSP\u2010GFP without disruption of oocyst egress and salivary gland invasion.<\/jats:p>","DOI":"10.1111\/mmi.15230","type":"journal-article","created":{"date-parts":[[2024,2,24]],"date-time":"2024-02-24T04:51:10Z","timestamp":1708750270000},"page":"565-577","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Overcoming the egress block of <i>Plasmodium<\/i> sporozoites expressing fluorescently tagged circumsporozoite protein"],"prefix":"10.1111","volume":"121","author":[{"given":"Carolina","family":"Thieleke\u2010Matos","sequence":"first","affiliation":[{"name":"Integrative Parasitology Center for Infectious Diseases, Heidelberg University Medical School  Heidelberg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-8316-9808","authenticated-orcid":false,"given":"Kevin","family":"Walz","sequence":"additional","affiliation":[{"name":"Integrative Parasitology Center for Infectious Diseases, Heidelberg University Medical School  Heidelberg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8332-6668","authenticated-orcid":false,"given":"Friedrich","family":"Frischknecht","sequence":"additional","affiliation":[{"name":"Integrative Parasitology Center for Infectious Diseases, Heidelberg University Medical School  Heidelberg Germany"},{"name":"German Center for Infection Research DZIF, Partner site Heidelberg  Heidelberg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5757-2750","authenticated-orcid":false,"given":"Mirko","family":"Singer","sequence":"additional","affiliation":[{"name":"Integrative Parasitology Center for Infectious Diseases, Heidelberg University Medical School  Heidelberg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2024,2,23]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2009.03.008"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20101488"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20040989"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2007.10.002"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pt.2015.04.010"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0113923"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462\u20105822.2004.00447.x"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1186\/1475\u20102875\u201011\u2010421"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.210971"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2209729119"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/385336a0"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/29.9.e45"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006396"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2019"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1111\/mmi.12528"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1111\/cmi.13321"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13059\u2010015\u20100811\u20101"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2019.10.030"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/21.7.1586"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462\u20105822.2005.00579.x"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.0010009"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.6985745"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467\u2010022\u201030988\u2010z"}],"container-title":["Molecular 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