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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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García‐Tuñón, I., Hernández‐Sánchez, M., Ordóñez, J. L., Alonso-Pérez, V., Quijada‐Álamo, M., Benito, R., Guerrero, C., Hernández‐Rivas, J. M., & Sánchez‐Martín, M. (2017) · Oncotarget · doi:10.18632/oncotarget.15215 · Cited by 41
McLemore, A. F., Hou, H., Meyer, B. S., Lam, N. B., Ward, G., Aldrich, A. L., Rodrigues, M. A., Vedder, A., Zhang, L., Padron, E., et al. (2022) · JCI Insight · doi:10.1172/jci.insight.159430 · Cited by 42
Current application of CRISPR/Cas9 gene-editing technique to eradication of HIV/AIDS
Huang, Z., Tomitaka, A., Raymond, A., & Nair, M. (2017) · Gene Therapy · doi:10.1038/gt.2017.35 · Cited by 43
Xu, X., Koivisto, O., Liu, C., Zhou, J., Miihkinen, M., Jacquemet, G., Wang, D., Rosenholm, J. M., Shu, Y., & Zhang, H. (2020) · Advanced Therapeutics · doi:10.1002/adtp.202000072 · Cited by 41
Ward, G., McGraw, K. L., Aghababazadeh, F. A., Meyer, B. S., McLemore, A. F., Vincelette, N. D., Lam, N. B., Aldrich, A. L., Ali, N. H. A., Padron, E., et al. (2021) · Blood Advances · doi:10.1182/bloodadvances.2020003475 · Cited by 42
A prolactin-dependent sexually dimorphic mechanism of migraine chronification
Ikegami, D., Navratilova, E., Yue, X., Moutal, A., Kopruszinski, C. M., Khanna, R., Patwardhan, A. M., Dodick, D. W., & Porreca, F. (2021) · Cephalalgia · doi:10.1177/03331024211039813 · Cited by 40
Matsuura-Suzuki, E., Shimazu, T., Takahashi, M., Kotoshiba, K., Suzuki, T., Kashiwagi, K., Sohtome, Y., Akakabe, M., Sodeoka, M., Dohmae, N., et al. (2022) · eLife · doi:10.7554/elife.72780 · Cited by 42
Shen, H. M., McHale, C. M., Smith, M. T., & Zhang, L. (2015) · Mutation Research/Reviews in Mutation Research · doi:10.1016/j.mrrev.2015.01.002 · Cited by 40
CRISPR‐Cas9 systems: versatile cancer modelling platforms and promising therapeutic strategies
Wen, W., Yuan, Z., Ma, S., Jiang, X., & Yuan, D. (2015) · International Journal of Cancer · doi:10.1002/ijc.29626 · Cited by 41
CRISPR/Cas9 editing of the genome for cancer modeling
Guernet, A. A., & Grumolato, L. (2017) · Methods · doi:10.1016/j.ymeth.2017.03.007 · Cited by 44
CRISPR to fix bad blood: a new tool in basic and clinical hematology
González-Romero, E., Martínez-Valiente, C., García-Ruiz, C., Vázquez‐Manrique, R. P., Cervera, J., & Sanjuán-Pla, A. (2019) · Haematologica · doi:10.3324/haematol.2018.211359 · Cited by 42
Lee, B., Lozano, R. J., & Dunbar, C. E. (2021) · Molecular Therapy · doi:10.1016/j.ymthe.2021.09.001 · Cited by 39
Genome-wide siRNA screen identifies UNC50 as a regulator of Shiga toxin 2 trafficking
Selyunin, A. S., Iles, L. R., Bartholomeusz, G. A., & Mukhopadhyay, S. (2017) · The Journal of Cell Biology · doi:10.1083/jcb.201704015 · Cited by 39
Curative Ex Vivo Hepatocyte-Directed Gene Editing in a Mouse Model of Hereditary Tyrosinemia Type 1
VanLith, C. J., Guthman, R. M., Nicolas, C. T., Allen, K. L., Du, Z., Joo, D. J., Nyberg, S. L., Lillegard, J. B., & Hickey, R. (2018) · Human Gene Therapy · doi:10.1089/hum.2017.252 · Cited by 39
Modelling acute myeloid leukemia (AML): What’s new? A transition from the classical to the modern
Dozzo, A., Galvin, A., Shin, J., Scalia, S., O’Driscoll, C. M., & Ryan, K. B. (2022) · Drug Delivery and Translational Research · doi:10.1007/s13346-022-01189-4 · Cited by 40
CRISPR-Cas Technology as a Revolutionary Genome Editing tool: Mechanisms and Biomedical Applications
Ebrahimi, S., Khosravi, M. A., Raz, A., Karimipoor, M., & Parvizi, P. (2023) · ̒Ulūm va funūn-i bastah/bandī · doi:10.61186/ibj.27.5.219 · Cited by 40
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Inactivation of Cancer Mutations Utilizing CRISPR/Cas9
Gebler, C., Lohoff, T., Paszkowski‐Rogacz, M., Mircetic, J., Chakraborty, D., Camgöz, A., Hamann, M. V., Theis, M., Thiede, C., & Buchholz, F. (2016) · JNCI Journal of the National Cancer Institute · doi:10.1093/jnci/djw183 · Cited by 38
Binding of NUFIP2 to Roquin promotes recognition and regulation of ICOS mRNA
Rehage, N., Davydova, E., Conrad, C., Behrens, G., Maiser, A., Stehklein, J. E., Brenner, S., Klein, J., Jeridi, A., Hoffmann, A., et al. (2018) · Nature Communications · doi:10.1038/s41467-017-02582-1 · Cited by 36
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Deubiquitinase OTUD5 is a positive regulator of mTORC1 and mTORC2 signaling pathways
Cho, J. H., Kim, K., Kim, S., Park, B.-O., Park, B.-O., Kim, J.-H., Kim, S., Kwon, M. J., Han, M., Lee, S. B., et al. (2020) · Cell Death and Differentiation · doi:10.1038/s41418-020-00649-z · Cited by 37
The PAF1 complex promotes 3′ processing of pervasive transcripts
Liu, X., Guo, Z., Han, J., Peng, B., Zhang, B., Li, H., Hu, X., David, C. J., & Chen, M. (2022) · Cell Reports · doi:10.1016/j.celrep.2022.110519 · Cited by 36
The retroviral vector family: something for everyone
Elsner, C., & Bohne, J. (2017) · Virus Genes · doi:10.1007/s11262-017-1489-0 · Cited by 36
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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