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Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing
Open accessAbstract
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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“Examples of LV delivery of CRISPR/Cas9 include modification of up to five genes using a single LV to deliver plasmid DNA encoding Cas9, sgRNA and a fluorescent marker to develop a mouse model of acute myeloid leukemia (Heckl et al., 2014)”
From Delivering CRISPR: a review of the challenges and approaches (2018) · cites this paper as Heckl et al., 2014
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“A great example is the mouse model of acute myeloid leukemia (AML) generated by modifying up to five genes in a single hematopoietic stem cell to mimic the genetic complexity of malignancy [57].”
From Viral Delivery Systems for CRISPR (2019) · cites this paper as [57]
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“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].”
From Non-viral delivery systems for CRISPR/Cas9-based genome editing: challenges and opportunities (2018) · cites this paper as [34]
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Metadata
- 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
- ORCID
- Venue
- Nature Biotechnology
- Published
- 2014-06-22
- Volume
- 32
- Issue
- 9
- Pages
- 941-946
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- PMC
- PMC4160386
- 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
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- 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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