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Updated: Apr 27, 2026
Lentiviral CRISPR/Cas9-Mediated Genome Editing for the Study of Hematopoietic Cells in Disease Models
Published on: October 3, 2019
Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing
Dirk Heckl1, Monika S Kowalczyk2, David Yudovich1
11] Division of Hematology, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA. [2].
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.
Insights
CRISPR-Cas9 genome editing enables rapid generation of complex mouse models for myeloid malignancy. This approach engineers multiple gene mutations in hematopoietic stem cells, better reflecting human cancer genetics.
Area of Science:
- Genetics
- Cancer Biology
- Molecular Biology
Background:
- Human malignancies often involve mutations in multiple driver genes.
- Conventional breeding methods struggle to create mouse models with such genetic complexity.
Purpose of the Study:
- To overcome limitations in creating genetically complex mouse cancer models.
- To utilize CRISPR-Cas9 genome editing for engineering in vivo cancer models.
Main Methods:
- Employing the CRISPR-Cas9 system with lentiviral vectors.
- Delivering combinations of small guide RNAs (sgRNAs) and Cas9 to mouse hematopoietic stem cells (HSCs).
- Modifying up to five genes simultaneously in HSCs.
Main Results:
- Successfully generated models of acute myeloid leukemia (AML) with cooperating mutations.
- Recapitulated combinations of mutations found in human AML patients.
- Demonstrated clonal outgrowth and myeloid malignancy in engineered mice.
Conclusions:
- Lentivirus-delivered CRISPR-Cas9 genome editing is effective for engineering complex in vivo cancer models.
- This method allows for the creation of models that better reflect the genetic complexity of human diseases.
- The approach is applicable to a broad array of cancer types.
