Purchase of high performance flow electroporation system for genome engineering facility

Proper diagnosis of genetic diseases is very important for patients and their families. Thanks to advances in genome sequencing, doctors are now able to list all the mutations present in a patient's genome - but the interpretation of these mutations is difficult. In this project, we will perform experiments to systematically study the effects of mutations in a number of disease-associated genes - including thousands of mutations that have not yet been observed in a patient, but are likely to be observed in the future.

We will study genes associated with blood calcium and glucose content, neurological disease, development of facial features in the embryo, and cancer. Studying mutations in these genes is a lengthy process that includes the transfer the DNA into cultured cells using an electric field, in a process called "electroporation". The equipment funded by the MRC will greatly enhance the efficiency of electroporation, which will increase the number of mutations we can study, as well as the accuracy of measurements. The knowledge of the effects of mutations will facilitate diagnosis, and it may also help select the appropriate drug depending on the type of mutation carried by the patient - a goal known as "precision medicine"

A key aim of research at the HGU is to understand the molecular basis of human development and genetic disease. A promising strategy, enabled by recent advances in synthetic biology and next generation sequencing, involves reverse genetic screens using large libraries of genetic variants. Examples include CRISPR/Cas9 screening, in which a guide RNA library is introduced into cells to disrupt genes one at a time, and deep mutational scanning (DMS), where all possible amino acid changes are introduced into the target gene. Such reverse genetic screens can be implemented in an ever-increasing range of cellular models to interrogate a growing number of phenotypes, with enormous potential to illuminate the genetic mechanisms of disease, to accelerate rate determining steps in genetic diagnosis, and to facilitate translational efforts to find new treatments for rare diseases.

Through DMS screens in mammalian cells, we will systematically determine the effect of mutations in hyper/hypocalcaemia and cancer (Kudla), insulin resistance syndromes (Semple), face development (Long), neurological disease (Gilbert); and the role of epimutations in cancer (Sproul). These projects are directly relevant to Prevention and Early Detection of Disease and Precision Medicine. By subjecting cellular mutation libraries to advanced cell phenotyping and small molecule and genetic screens, and by developing new tools and strategies for targeted protein degradation in vivo (Wood), we will both identify responses of mutant proteins to available drugs, and will facilitate the targetting of hitherto undruggable proteins. The functionality of the MaxCyte ExPERT GTx system will permit efficient implementation of DMS pipelines and their articulation with cellular phenotyping and advanced therapy pipelines in a manner ideally suited to creating a Rare Disease Hub in Edinburgh accessible to the UK Rare Disease community.