Genome editing to model and reverse a prevalent mutation associated with myeloproliferative neoplasms.

摘要: Myeloproliferative neoplasms (MPNs) cause the over-production of blood cells such as erythrocytes (polycythemia vera) or platelets (essential thrombocytosis). JAK2 V617F is most prevalent somatic mutation in many MPNs, but previous modeling this mice relied on transgenic overexpression and resulted diverse phenotypes that were some cases attributed to expression level. CRISPR-Cas9 engineering offers new possibilities model potentially cure genetically encoded disorders via precise modification endogenous locus primary cells. Here we develop "scarless" Cas9-based reagents create reverse an immortalized human erythroid progenitor cell line (HUDEP-2), CD34+ adult hematopoietic stem (HSPCs), immunophenotypic long-term (LT-HSCs). We find no overt vitro increase proliferation associated with allele, co-culture wild type unmasks a competitive growth advantage provided by mutation. Acquisition allele also promotes terminal differentiation progenitors, even absence cytokine signaling. Taken together, these data are consistent gradually progressive manifestation MPNs reveals endogenously acquired mutations may yield more subtle compared models.