Alternative CRISPR-Cas Enzymes to Advance Gene Drive Strategies in Malaria Vectors

This PhD opportunity is being offered as part of the LSTM and Lancaster University Doctoral Training Partnership. Find out more about the studentships and how to apply. 

Abstract Malaria remains a significant public health challenge, with recent increases in global cases highlighting the need for new and novel vector control technologies. Genetic vector control is one such technique which uses CRISPR to introduce gene edits with the aim of either modifying or reducing the target population. Our group uses a technique known as gene drive to bias the inheritance of these edited genetic elements to enable more rapid spread of the desired gene edit and ultimately a faster modification or reduction in the target population. The majority of CRISPR gene editing approaches rely on the use of Cas9, an endonuclease which acts as molecular scissors cutting the DNA at precise locations on the genome. These “molecular scissors” are integral to CRISPR systems as they allow precise insertions or deletions in specific target regions. More recent identification of other Cas enzymes provides opportunity to expand the CRISPR toolkit beyond the use of Cas9. Using these alternative enzymes may enable editing in previously unavailable genomic locations, more efficient transformation, reduction in problematic off target effects and if used in combination with Cas9 reduce development of resistance to gene drives. This project will therefore explore alternative Cas9 architectures in the design of genetic control strategies for malaria vectors including Anopheles gambiae and funestus. This will include identification of suitable alternatives to Cas9 and their integration into the design of new genetic constructs, their integration into the mosquito genome and characterisation of their performance. These alternative Cas enzymes will be used to introduce edits at selected genomic target sites to assess their efficiency in contributing to the development of more efficient genetic control.
Where does this project lie in the translational pathway? T1 - Basic Research,T3 - Evidence into Practice
Methodological Aspects
  • Bioinformatics skills
  • Molecular biology and cloning
  • CRISPR genome editing
  • Embryo microinjection and generation of transgenic mosquito lines
  • Characterisation of transgenic lines.
  • Cross and cage based population simulations.
  • Modelling gene drives
Expected Outputs The project will produce high quality REF returnable 3*/4* publications and will provide the evidence base for large scale research council, philanthropic (e.g. OP and Gates) and industry funding in a global priority area. the development of an international profile in mosquito genetics and gene drive research; transferable personal skills to industry and/or academia.
Training Opportunities
  • CRISPR genome editing
  • DNA cloning and molecular biology techniques
  • Execution of large cage studies
  • Bioinformatics
  • Insect husbandry
  • Professional development course
  • Data analysis and presentation
  • Publication and report writing
Skills Required
  • A good general understanding of genetics
  • An aptitude for molecular biology, genetics and vector control.
  • Bioinformatics experience would be beneficial but is not essential.
Subject Areas Malaria and other Vector Borne Diseases
Key Publications associated with this project

Abudayyeh, O.O., Gootenberg, J.S., Essletzbichler, P., Han, S., Joung, J., Belanto, J.J., Verdine, V., Cox, D.B., Kellner, M.J., Regev, A. and Lander, E.S., 2017. RNA targeting with CRISPR–Cas13. Nature, 550(7675), pp.280-284.

Yan, F., Wang, W. and Zhang, J., 2019. CRISPR-Cas12 and Cas13: the lesser known siblings of CRISPR-Cas9. Cell biology and toxicology, 35(6), pp.489-492.

Collier, T.C., Lee, Y., Mathias, D.K. and López Del Amo, V., 2024. CRISPR-Cas9 and Cas12a target site richness reflects genomic diversity in natural populations of Anopheles gambiae and Aedes aegypti mosquitoes. BMC genomics, 25(1), p.700.