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Updated: Sep 10, 2025
Studying Cryptosporidium Infection in 3D Tissue-derived Human Organoid Culture Systems by Microinjection
Published on: September 14, 2019
Targeted CRISPR screens reveal genes essential for Cryptosporidium survival in the host intestine
Lucy C Watson1, Katarzyna A Sala1, Netanya Bernitz1
1Cryptosporidiosis Laboratory, The Francis Crick Institute, London, UK.
Insights
A new CRISPR screening tool helps identify essential Cryptosporidium genes. Researchers found gene Cp23 is vital for parasite survival and motility, crucial for reinfection.
Area of Science:
- * Infectious Diseases
- * Parasitology
- * Molecular Biology
Background:
- * Cryptosporidium is a major cause of childhood diarrhea and malnutrition, with limited treatment options.
- * Developing new therapies is hindered by a lack of understanding of essential parasite genes.
- * Current treatment options for Cryptosporidium infections are limited, necessitating novel therapeutic strategies.
Purpose of the Study:
- * To develop and apply a CRISPR-based screening method for rapid genetic analysis of Cryptosporidium.
- * To identify essential genes for Cryptosporidium survival and virulence in vivo.
- * To investigate the role of the pyrimidine salvage pathway and vaccine candidates in parasite biology.
Main Methods:
- * Iterative improvement of genetic manipulation tools for Cryptosporidium.
- * Development of a targeted CRISPR-based screening assay for gene essentiality.
- * Application of inducible knockout to assess gene function in vivo.
Main Results:
- * A novel CRISPR screening method was established for efficient gene essentiality assessment.
- * The parasite gene Cp23 was identified as essential for Cryptosporidium survival in vivo.
- * Cp23-deficient parasites exhibit defects in gliding motility, impacting reinfection capability.
Conclusions:
- * The developed CRISPR screening platform accelerates the study of Cryptosporidium infection biology.
- * Identification of Cp23 as essential provides a new target for therapeutic intervention.
- * This work lays the groundwork for developing novel treatments against Cryptosporidium infections.
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.
