Universal repair of ataxia telangiectasia mutations by CRISPR genome editing

Ataxia telangiectasia (A-T) is a severe autosomal recessive genetic disease caused by mutations in the ATM gene, which lead to cerebellar degeneration, cancer susceptibility and immunodeficiency. ATM is a very large gene spanning 66 exons, constitutively expressed in all cells. ATM encodes the apical ATM kinase, which plays a central role in the cellular response to DNA double strand breaks (DSBs), apoptosis and cell-cycle checkpoint control. No effective treatment is currently available for A-T, but correction of the mutated gene through CRISPR/Cas genome editing is feasible. However, mutations in ATM are spread across the whole gene, with no evidence of hot-spots, complicating the development of gene correction approaches of wide applicability. We are interested in the design of such universal strategies to correct ATM, based on the replacement of large segments of the gene through genome editing. We have designed and built repair templates to achieve this correction strategy, and identified and tested suitable CRISPR target sites within ATM. This project will now test and optimise such CRISPR/Cas genome editing approaches to correct multiple mutations in ATM, initially in suitable cell lines and eventually in human haematopoietic stem cells. Clinical correction of haematopoietic stem cells would be expected to alleviate the immunodeficiency and lymphoid malignancy risk observed in people affected by A-T. The project involves the use of advanced genetic engineering, viral vectors (integration-deficient lentiviral vectors and/or adeno-associated viral vectors), cell culture and multiple biochemical and molecular downstream analyses. The work will be done in our Royal Holloway University of London laboratory (http://AGCTlab.org), which is focused on the development of new gene and stem cell therapies for rare and common diseases, and may also involve colleagues in other institutions.

Candidates are expected to have a BSc (Upper second class or above) in a relevant discipline, knowledge and experience of molecular cell biology and genome editing. Experience with viral vectors would be an advantage

To apply follow link and instructions at https://www.royalholloway.ac.uk/studying-here/applying/postgraduate/how-to-apply/. Please indicate supervisor’s last name "Yáñez-Muñoz" and project title in your application. Application deadline 12 March 2023