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Following ingestion during blood feeding, parasites reproduce sexually and traverse the mosquito midgut epithelium to differentiate into oocysts on the basal lamina, where they undergo prolonged development, ultimately giving rise to thousands of sporozoites capable of infecting a new human host. Oocyst formation represents a severe population bottleneck, resulting in the lowest parasite numbers observed across the parasite life cycle. Given its extended duration and pronounced numerical vulnerability, it is striking that the oocyst remains one of the least explored stages of Plasmodium development. Major gaps persist in our understanding of the molecular and cellular processes governing oocyst growth and differentiation, including transcriptional and epigenetic regulation, nutrient acquisition and metabolic remodelling, cell cycle control and interactions with the mosquito immune system and physiology. Recent technological advances and renewed interest in mosquito-stage biology provide an opportunity to dissect these processes at unprecedented resolution. In this review, we synthesize knowledge of oocyst biology, highlight key unresolved questions and discuss how deeper insight into this critical stage could inform the development of next-generation transmission-blocking strategies and accelerate progress towards malaria elimination.<\/jats:p>","DOI":"10.1098\/rsob.260009","type":"journal-article","created":{"date-parts":[[2026,4,22]],"date-time":"2026-04-22T00:08:23Z","timestamp":1776816503000},"source":"Crossref","is-referenced-by-count":2,"title":["Oocyst: knowns and unknowns about the lengthiest life stage of the malaria parasite"],"prefix":"10.1098","volume":"16","author":[{"given":"Alexander J.","family":"Bailey","sequence":"first","affiliation":[{"name":"Imperial College London 1 , ,","place":["London, UK"],"department":["Department of Life Sciences"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dina","family":"Vlachou","sequence":"additional","affiliation":[{"name":"Imperial College London 1 , ,","place":["London, UK"],"department":["Department of Life Sciences"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3323-1687","authenticated-orcid":false,"given":"George K.","family":"Christophides","sequence":"additional","affiliation":[{"name":"Imperial College London 1 , ,","place":["London, UK"],"department":["Department of Life Sciences"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"175","published-online":{"date-parts":[[2026,4,22]]},"reference":[{"key":"2026042120081815800_R1","doi-asserted-by":"crossref","DOI":"10.1371\/journal.ppat.1012965","article-title":"Advances in the dissection of Anopheles\u2013Plasmodium interactions","volume":"21","author":"Saab","year":"2025","journal-title":"PLoS Pathog."},{"key":"2026042120081815800_R2","doi-asserted-by":"crossref","first-page":"644","DOI":"10.1590\/0074-0276130597","article-title":"The Plasmodium bottleneck: malaria parasite losses in the mosquito vector","volume":"109","author":"Smith","year":"2014","journal-title":"Mem. 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