Using human disease and genome engineering to dissect dynamic genome organisation at the nuclear lamina in vivo during development

The specialisation of an embryo’s cells into the many cell types of the body is achieved by precisely activating or inactivating specific genes at the appropriate time and place. To achieve this precision, genes are isolated from the noise of the wider genome by locally folding with nearby DNA to form discrete structures called topologically-associated domains (TADs). When this fails because of removal or rearrangement of TAD boundaries by mutation there will be inappropriate gene activity and disease. During development some TADs also attach and detach from the nuclear lamina at the nucleus’s outer edge, turning associated genes off. However, it is unknown if disease-causing TAD-rearranging mutations also disrupt lamina attachment, impairing its control of gene activity.

I will measure altered lamina attachment in developing mouse limbs that have TAD-rearranging mutations and monitor how this pathogenically affects gene activity and limb formation. I will also develop methods to specifically observe lamina-attached TAD structures and measure how disease-causing mutations disrupt them.

This research will unravel lamina attachment’s purpose in development, allowing the effects of mutations that alter the genome’s 3D organisation to be predicted in patients.