Cited by 557CRISPR and Genetic EngineeringAcute Myeloid Leukemia ResearchChromatin Remodeling and Cancer

Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing

Open access
Heckl, Dirk, Kowalczyk, Monika S, Yudovich, David, Belizaire, Roger, Puram, Rishi V, McConkey, Marie E, Thielke, Anne, Aster, Jon C +2 more·2014·Nature Biotechnology

Abstract

Genome sequencing studies have shown that human malignancies often bear mutations in four or more driver genes, but it is difficult to recapitulate this degree of genetic complexity in mouse models using conventional breeding. Here we use the CRISPR-Cas9 system of genome editing to overcome this limitation. By delivering combinations of small guide RNAs (sgRNAs) and Cas9 with a lentiviral vector, we modified up to five genes in a single mouse hematopoietic stem cell (HSC), leading to clonal outgrowth and myeloid malignancy. We thereby generated models of acute myeloid leukemia (AML) with cooperating mutations in genes encoding epigenetic modifiers, transcription factors and mediators of cytokine signaling, recapitulating the combinations of mutations observed in patients. Our results suggest that lentivirus-delivered sgRNA:Cas9 genome editing should be useful to engineer a broad array of in vivo cancer models that better reflect the complexity of human disease.

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  • Viral vectors have been extensively applied to deliver CRISPR cassettes in vitro and in vivo largely owing to their high efficiency in gene delivery and long-term stable transgene expression.

    “In recent years, viral vectors have been extensively applied to deliver CRISPR cassettes in vitro and in vivo largely owing to their high efficiency in gene delivery and long-term stable transgene expression [34, 58, 65].”

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Authors
Heckl, D., Kowalczyk, M. S., Yudovich, D., Belizaire, R., Puram, R. V., McConkey, M. E., Thielke, A., Aster, J. C., Regev, A., & Ebert, B. L.
Affiliations
  • Brigham and Women's Hospital (US)
  • Broad Institute (US)
Corresponding author
Ebert, Benjamin L
Venue
Nature Biotechnology
Published
2014-06-22
Volume
32
Issue
9
Pages
941-946
ISSN
  • 0733-222X (ISSN-L)
  • 1087-0156
  • 1546-1696
OpenAlex
W1575193685
Cited by
555
References
27
Citations by year
2026: 14 · 2025: 29 · 2024: 28 · 2023: 58 · 2022: 58 · 2021: 58 · 2020: 61 · 2019: 51 · 2018: 54 · 2017: 61 · 2016: 38 · 2015: 39 · 2014: 5
FWCI
19.53
Type
journal article
Language
en
Open access
Locations
  • Nature Biotechnology (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • Digital Access to Scholarship at Harvard (DASH) (Harvard University) (submittedVersion, cc-by)
  • DSpace@MIT (Massachusetts Institute of Technology) (submittedVersion, cc-by-nc-sa)
Funders
  • National Heart, Lung, and Blood Institute (R01HL082945)
  • National Human Genome Research Institute (P50HG006193)
  • National Cancer Institute (P01CA108631, P01CA066996)
  • National Institute of General Medical Sciences (T32GM007753)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Health Sciences › Medicine › Hematology › Acute Myeloid Leukemia Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Chromatin Remodeling and Cancer
Keywords
myeloid neoplasms, acute myeloid leukemia, mouse tumor model, CRISPR/Cas9 genome editing, lentiviral delivery, hematopoietic stem cells, clonal outgrowth, driver mutations, epigenetic regulators, transcription factors, cytokine signaling, cooperating mutations
MeSH terms
Animals, Disease Models, Animal, Bone Marrow Neoplasms, Mice, Clustered Regularly Interspaced Short Palindromic Repeats

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