Dual epigenetic and genetic editing of primary human haematopoietic stem and progenitor cells

Event details

Abstract: The epigenome is perturbed in haematological malignancies including localized DNA hypermethylation at the gene promoter CpG islands. Understanding the impact of these hypermethylation changes on normal cell function can be investigated by using a modified form of CRISPR/Cas9 which targets DNA methylation as opposed to creating mutations. In this study, we show for the first time that de novo DNA methylation can be targeted using CRISPR/Cas9 in human hematopoietic stem and progenitor cells, and we multiplexed DNA methylation editing at cell cycle gene promoters INK4b (p15) and ARF (p14). Bulk and single myeloid colony analysis after 14 days of differentiation show that hypermethylation is maintained during myeloid differentiation, inhibiting gene expression.

Since genetic mutations also play a key role in myeloid cancer development, we further adapted the method to edit the epigenome and genome in the same haematopoietic cell. By combining epigenetic and genetic editing, we can recapitulate disease-relevant hits using CRISPR/dCas9 to help us study the cellular processes that may underlie myeloid cancer development.