Priority Research Papers:

ASXL1 mutation correction by CRISPR/Cas9 restores gene function in leukemia cells and increases survival in mouse xenografts

Simona Valletta, Hamid Dolatshad, Matthias Bartenstein, Bon Ham Yip, Erica Bello, Shanisha Gordon, Yiting Yu, Jacqueline Shaw, Swagata Roy, Laura Scifo, Anna Schuh, Andrea Pellagatti, Tudor A. Fulga, Amit Verma and Jacqueline Boultwood _

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Oncotarget. 2015; 6:44061-44071. https://doi.org/10.18632/oncotarget.6392

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Simona Valletta1, Hamid Dolatshad1, Matthias Bartenstein2, Bon Ham Yip1, Erica Bello1, Shanisha Gordon2, Yiting Yu2, Jacqueline Shaw1, Swagata Roy1, Laura Scifo1, Anna Schuh3, Andrea Pellagatti1, Tudor A. Fulga4, Amit Verma2, Jacqueline Boultwood1

1Bloodwise Molecular Haematology Unit, Nuffield Division of Clinical Laboratory Sciences, Radcliffe Department of Medicine, University of Oxford and BRC Blood Theme, NIHR Oxford Biomedical Centre, Oxford University Hospital, Oxford, UK

2Albert Einstein College of Medicine, Bronx, NY, USA

3NIHR Biomedical Research Centre, University of Oxford, Oxford, UK

4Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of Oxford, John Radcliffe Hospital, Oxford, UK

Correspondence to:

Jacqueline Boultwood, e-mail: [email protected]

Keywords: ASXL1, CRISPR, chronic myeloid leukemia, mutation correction, tumor suppressor

Received: July 22, 2015     Accepted: November 08, 2015     Published: November 26, 2015


Recurrent somatic mutations of the epigenetic modifier and tumor suppressor ASXL1 are common in myeloid malignancies, including chronic myeloid leukemia (CML), and are associated with poor clinical outcome. CRISPR/Cas9 has recently emerged as a powerful and versatile genome editing tool for genome engineering in various species. We have used the CRISPR/Cas9 system to correct the ASXL1 homozygous nonsense mutation present in the CML cell line KBM5, which lacks ASXL1 protein expression. CRISPR/Cas9-mediated ASXL1 homozygous correction resulted in protein re-expression with restored normal function, including down-regulation of Polycomb repressive complex 2 target genes. Significantly reduced cell growth and increased myeloid differentiation were observed in ASXL1 mutation-corrected cells, providing new insights into the role of ASXL1 in human myeloid cell differentiation. Mice xenografted with mutation-corrected KBM5 cells showed significantly longer survival than uncorrected xenografts. These results show that the sole correction of a driver mutation in leukemia cells increases survival in vivo in mice. This study provides proof-of-concept for driver gene mutation correction via CRISPR/Cas9 technology in human leukemia cells and presents a strategy to illuminate the impact of oncogenic mutations on cellular function and survival.

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