Klin Padiatr 2015; 227 - A1
DOI: 10.1055/s-0035-1550233

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

J Reimer 1, S Knoess 1, M Labuhn 1, JH Klusmann 1, D Heckl 1
  • 1Ped. Hematology & Oncology, Hannover Medical School

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