Improved lentiviral CRISPR-Cas9 vectors for the generation of leukemogenic chromosomal translocations

Introduction: AML subgroups are cytogenetically defined by recurrent chromosomal translocation (CTs), which are causative for leukemic transformation. CRISPR-Cas9-based genome editing emerged as a powerful tool for modelling CTs. However, broad application to primary hematopoietic stem and progenitor cells (HSPCs) was limited due to inefficient delivery tools. Methods: Here, we generated and evaluated improved lentiviral CRISPR-Cas9 vectors (lentiCRISPR-CT2.0) with new architecture and two sgRNA expression cassettes. Results: lentiCRISPR-CT2.0 vectors generated significantly higher titers, allowing efficient tra…