Exploring gene functions in an obligate intracellular parasite Theileria using CRISPR-Cas9 technologies

Exploring gene functions in an obligate intracellular parasite Theileria using CRISPR-Cas9 technologies

Internship Description

Background: Theileria is a unique and remarkable apicomplexan parasite capable of transforming its host leukocyte into a disseminating cancer-like tumour and it is the only known example of natural reversible transformation of mammalian host leukocytes by an eukaryotic parasite. T. annulata is the causative agent of the cattle disease called tropical theileriosis, which is of major economic importance in countries in Northern Africa, The Middle East and Asia. Importantly, the tumor-like phenotype is reversed upon drug-induced parasite death. Moreover, virulent macrophages can be attenuated by multiple in vitro passages, and upon attenuation, they lose both adhesion and invasiveness. Transfection systems are available for several important species of protozoa, including the apicomplexan parasites Toxoplasma gondii, Eimeria tenella and Plasmodium spp. However, such approaches are not available for Theileria spp.

Objectives: In this study, we aim to test the feasibility of new delivery systems to create genetic tools for the study of T. annulata genes. The CRISPRCas9 system is a powerful tool for gene editing in various cells and organisms, but delivery of CRISPRCas9 plasmids into cells or tissues is challenging, because the vectors encoding both Cas9 and sgRNA are handicapped by both a strong negative charge and a large size.

Methodology: In order to achieve our objective, we propose to develop nanoparticles to deliver the CRISPR genome-editing system into T. annulata-infected leukocytes and specifically modify expressed parasite genes. This study will be performed in active collaboration with KAUST experts in the nanomaterials field.​


This study will fill a technology gap in Theileria research by enabling systematic and genome-wide functional studies of parasite gene functions, and facilitating assessment of many aspects of the biology of this unique host-apicomplexan parasite interaction. Moreover, defined vaccines created by genetically manipulating parasite genes and validation of antigens will become a possibility.

Faculty Name

Arnab Pain

Field of Study

Microbiology, Molecular Biology, Cell biology, Parasitology