CRISPR/Cas9 Genome Editing to Model AD Risk Variants with Isogenic Pluripotent Stem Cells

Theme 4.3

In this work package, we plan to combine iPSc and genome editing technology to further our understanding of Alzheimer Disease (AD) genetics and pathogenesis. We will use patient-derived iPSCs to model known causal variants in AD (for example rs59335482 in BIN1 and rs75932628 in TREM2.

 

Using the latest CRISPR/Cas9 technology, we will introduce precise modifications into the genome of our patient-derived iPSCs. These modifications will ‘correct’ the putative risk alleles present in the AD patient line by replacing them with wild type alleles. Following genome editing, we will differentiate both unmodified and modified iPSC lines into disease-relevant cell types, such as cortical neurons, microglia and peripheral leucocytes.  Derived from the same patient, these cell lines will have an identical genetic background, which differ only in the presence or absence of a specific risk allele. These powerful models will allow us to determine the impact of risk variants on AD in a highly controlled manner.

Outputs of this work package will be the Creation of patient-derived isogenic human disease models by:

  • Editing the genome of patient-derived iPSCs to create isogenic cell lines
  • Differentiation of iPSCs into disease-relevant cell types such as cortical   neurons, microglia and peripheral leucocytes.

Risk genes to be investigated include BIN1, TREM2, PICALM and CD2AP.

 

Details of Work Package Lead
Prof Julie Williams - Cardiff University