CRISPR gene editing of murine blood stem and progenitor cells induces MLL-AF9 chromosomal translocation and MLL-AF9 leukaemogenesis

Sarrou, E. , Richmond, L., Carmody, R. J. , Gibson, B. and Keeshan, K. (2020) CRISPR gene editing of murine blood stem and progenitor cells induces MLL-AF9 chromosomal translocation and MLL-AF9 leukaemogenesis. International Journal of Molecular Sciences, 21(12), 4266. (doi: 10.3390/ijms21124266) (PMID:32549410) (PMCID:PMC7352880)

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Abstract

Chromosomal rearrangements of the mixed lineage leukaemia (MLL, also known as KMT2A) gene on chromosome 11q23 are amongst the most common genetic abnormalities observed in human acute leukaemias. MLL rearrangements (MLLr) are the most common cytogenetic abnormalities in infant and childhood acute myeloid leukaemia (AML) and acute lymphocytic leukaemia (ALL) and do not normally acquire secondary mutations compared to other leukaemias. To model these leukaemias, we have used clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 gene editing to induce MLL-AF9 (MA9) chromosomal rearrangements in murine hematopoietic stem and progenitor cell lines and primary cells. By utilizing a dual-single guide RNA (sgRNA) approach targeting the breakpoint cluster region of murine Mll and Af9 equivalent to that in human MA9 rearrangements, we show efficient de novo generation of MA9 fusion product at the DNA and RNA levels in the bulk population. The leukaemic features of MA9-induced disease were observed including increased clonogenicity, enrichment of c-Kit-positive leukaemic stem cells and increased MA9 target gene expression. This approach provided a rapid and reliable means of de novo generation of Mll-Af9 genetic rearrangements in murine haematopoietic stem and progenitor cells (HSPCs), using CRISPR/Cas9 technology to produce a cellular model of MA9 leukaemias which faithfully reproduces many features of the human disease in vitro.

Item Type:Articles
Additional Information:This research was funded by Kay Kendall Leukaemia Fund: NA, The Howat Foundation: NA, Cancer Research UK Glasgow Centre: C596/A18076, Cancer Research UK Beatson Institute: C596/A17196, Yorkhill Children’s Leukaemia Research Fund: NA, and Children with Cancer UK: NA.
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Sarrou, Evgenia and Gibson, Professor Brenda and Carmody, Dr Ruaidhri and Keeshan, Dr Karen and Richmond, Dr Laura
Authors: Sarrou, E., Richmond, L., Carmody, R. J., Gibson, B., and Keeshan, K.
College/School:College of Medical Veterinary and Life Sciences > Institute of Cancer Sciences
College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:International Journal of Molecular Sciences
Publisher:MDPI
ISSN:1661-6596
ISSN (Online):1422-0067
Published Online:15 June 2020
Copyright Holders:Copyright © 2020 by the authors.
First Published:First published in International Journal of Molecular Sciences 21(12):4266
Publisher Policy:Reproduced under a Creative Commons license

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
190874CR-UK Centre renewalKaren VousdenCancer Research UK (CRUK)C596/A18076Institute of Cancer Sciences