Targeted CRISPR screens reveal genes essential for Cryptosporidium survival in the host intestine
Abstract The Cryptosporidium parasite is one of the leading causes of diarrheal morbidity and mortality in children, and adolescent infections are associated with chronic malnutrition. There are no vaccines available for protection and only one drug approved for treatment that has limited efficacy....
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-08-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-63012-1 |
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| author | Lucy C. Watson Katarzyna A. Sala Netanya Bernitz Lotta Baumgärtel Mitchell A. Pallett N. Bishara Marzook Lorian Cobra Straker Duo Peng Lucy Collinson Adam Sateriale |
| author_facet | Lucy C. Watson Katarzyna A. Sala Netanya Bernitz Lotta Baumgärtel Mitchell A. Pallett N. Bishara Marzook Lorian Cobra Straker Duo Peng Lucy Collinson Adam Sateriale |
| author_sort | Lucy C. Watson |
| collection | DOAJ |
| description | Abstract The Cryptosporidium parasite is one of the leading causes of diarrheal morbidity and mortality in children, and adolescent infections are associated with chronic malnutrition. There are no vaccines available for protection and only one drug approved for treatment that has limited efficacy. A major barrier to developing new therapeutics is a lack of foundational knowledge of Cryptosporidium biology, including which parasite genes are essential for survival and virulence. Here, we iteratively improve the tools for genetically manipulating Cryptosporidium and develop a targeted CRISPR-based screening method to rapidly assess how the loss of individual parasite genes influence survival in vivo. Using this method, we examine the parasite’s pyrimidine salvage pathway and a set of leading Cryptosporidium vaccine candidates. From this latter group, using inducible knockout, we determined the parasite gene known as Cp23 to be essential for survival in vivo. Parasites deficient in Cp23 were able to replicate within and emerge from infected epithelial cells, yet unable to initiate gliding motility which is required for the reinfection of neighbouring cells. The targeted screening method presented here is highly versatile and will enable researchers to more rapidly expand the knowledge base for Cryptosporidium infection biology, paving the way for new therapeutics. |
| format | Article |
| id | doaj-art-fd6e0d4e5b3b467a8f371770160d48a6 |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-fd6e0d4e5b3b467a8f371770160d48a62025-08-24T11:37:14ZengNature PortfolioNature Communications2041-17232025-08-0116111310.1038/s41467-025-63012-1Targeted CRISPR screens reveal genes essential for Cryptosporidium survival in the host intestineLucy C. Watson0Katarzyna A. Sala1Netanya Bernitz2Lotta Baumgärtel3Mitchell A. Pallett4N. Bishara Marzook5Lorian Cobra Straker6Duo Peng7Lucy Collinson8Adam Sateriale9Cryptosporidiosis Laboratory, The Francis Crick InstituteCryptosporidiosis Laboratory, The Francis Crick InstituteCryptosporidiosis Laboratory, The Francis Crick InstituteCryptosporidiosis Laboratory, The Francis Crick InstituteCryptosporidiosis Laboratory, The Francis Crick InstituteCryptosporidiosis Laboratory, The Francis Crick InstituteElectron Microscopy Science Technology Platform, The Francis Crick InstituteChan Zuckerberg BiohubElectron Microscopy Science Technology Platform, The Francis Crick InstituteCryptosporidiosis Laboratory, The Francis Crick InstituteAbstract The Cryptosporidium parasite is one of the leading causes of diarrheal morbidity and mortality in children, and adolescent infections are associated with chronic malnutrition. There are no vaccines available for protection and only one drug approved for treatment that has limited efficacy. A major barrier to developing new therapeutics is a lack of foundational knowledge of Cryptosporidium biology, including which parasite genes are essential for survival and virulence. Here, we iteratively improve the tools for genetically manipulating Cryptosporidium and develop a targeted CRISPR-based screening method to rapidly assess how the loss of individual parasite genes influence survival in vivo. Using this method, we examine the parasite’s pyrimidine salvage pathway and a set of leading Cryptosporidium vaccine candidates. From this latter group, using inducible knockout, we determined the parasite gene known as Cp23 to be essential for survival in vivo. Parasites deficient in Cp23 were able to replicate within and emerge from infected epithelial cells, yet unable to initiate gliding motility which is required for the reinfection of neighbouring cells. The targeted screening method presented here is highly versatile and will enable researchers to more rapidly expand the knowledge base for Cryptosporidium infection biology, paving the way for new therapeutics.https://doi.org/10.1038/s41467-025-63012-1 |
| spellingShingle | Lucy C. Watson Katarzyna A. Sala Netanya Bernitz Lotta Baumgärtel Mitchell A. Pallett N. Bishara Marzook Lorian Cobra Straker Duo Peng Lucy Collinson Adam Sateriale Targeted CRISPR screens reveal genes essential for Cryptosporidium survival in the host intestine Nature Communications |
| title | Targeted CRISPR screens reveal genes essential for Cryptosporidium survival in the host intestine |
| title_full | Targeted CRISPR screens reveal genes essential for Cryptosporidium survival in the host intestine |
| title_fullStr | Targeted CRISPR screens reveal genes essential for Cryptosporidium survival in the host intestine |
| title_full_unstemmed | Targeted CRISPR screens reveal genes essential for Cryptosporidium survival in the host intestine |
| title_short | Targeted CRISPR screens reveal genes essential for Cryptosporidium survival in the host intestine |
| title_sort | targeted crispr screens reveal genes essential for cryptosporidium survival in the host intestine |
| url | https://doi.org/10.1038/s41467-025-63012-1 |
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