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DOI: 10.1055/s-0035-1550233
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 transduction of primary HSPCs. Dual sgRNA expression resulted in CTs t(9;11) and t(11;19), leading to expression of MLL-AF9 and MLL-ENL fusion transcripts.
Conclusions:
Second generation lentiCRISPR vectors show increased efficiency in modeling leukemogenic CTs in primary HSPCs, which will provide previously not accessible insights into the pathogenesis.
*contributed equally