Conditional knockout of essential genes in the malaria parasite

Key information

Application close date
07 February 2023, 11:59 GMT
Hours per week
36 (full time)
Application guidance
Posted 22 December 2022

Research topics

Biochemistry & Proteomics Infectious Disease
Background texture taken from the lab imagery.

This is a summer student position supervised by Abhinay Ramaprasad from Mike Blackman's lab. 

Introduction to the Science

Malaria is one of the oldest recorded diseases affecting mankind. It continues to exert a heavy toll on public health, claiming the lives of around half a million people each year.

Malaria is caused by a parasite called Plasmodium that invades our red blood cells (RBC). Once inside the RBC, the parasite grows and multiplies to form daughter parasites. The daughter parasites then break out of the RBC and go on to invade fresh RBCs. This replication cycle is essential for the parasite to survive and maintain its infection in the blood. The parasite genome contains around 5,000 genes, one-third of which are essential for optimal blood stage growth. In the Malaria Biochemistry Lab, we use conditional gene knockout approaches to study the role of different enzymes that the parasite needs to progress through its blood stage life cycle.

 

About the Project

We have a “hit list” of essential parasite enzymes that we want to conditionally knock out in the parasite. To do this, we first modify a target gene in the parasite’s genome by CRISPR/Cas9 gene editing. Next, we induce the deletion of the target gene (gene knockout) at a particular point in its life cycle. We then study how the deletion affects the genetically modified parasite. This will help us identify and understand the function of the enzyme in the parasite.

The candidate will help in the crucial first stage of this workflow – creating DNA constructs for genetic modification in the parasite. This will involve

  1. Designing the strategy for conditional knockout of a gene in the malaria parasite.
  2. Building CRISPR/Cas9 plasmids using several molecular cloning techniques. The candidate will learn to use PCR, seamless cloning, bacterial transformation and how to analyse sequencing data.
  3. The candidate will also be able to watch and learn techniques related to Plasmodium parasite culturing, transfection and studying its replication cycle.

 

About You

This project will suit a candidate with an interest in infectious diseases and who would like to learn about malaria parasites and their fascinating biology!

 

References 

1.         Knuepfer, E., Napiorkowska, M., van Ooij, C. and Holder, A.A. (2017)

            Generating conditional gene knockouts in Plasmodium - a toolkit to produce stable DiCre recombinase-expressing parasite lines using CRISPR/Cas9.

            Scientific Reports 7: 3881. PubMed abstract

2.         Ramaprasad, A., Burda, P.C., Calvani, E., Sait, A., Palma-Duran, S.A., Withers-Martinez, C., . . . Blackman, M.J. (2022)

            Preprint: A choline-releasing glycerophosphodiesterase essential for phosphatidylcholine biosynthesis and blood stage development in the malaria parasite.

            Available at: bioRxiv. https://www.biorxiv.org/content/10.1101/2022.06.14.496138