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Engineering Oncogenic Heterozygous Gain-of-Function Mutations in Human Hematopoietic Stem and Progenitor Cells
This content is Free Access.
1Division of Hematology, Department of Internal Medicine,Medical University of Graz, 2Division of Hematology, Stanford Cancer Institute, Stanford Institute for Stem Cell Biology and Regenerative Medicine,Stanford University School of Medicine, 3Department of Blood Group Serology and Transfusion Medicine,Medical University of Graz
Capitoli
- 00:05Introduction
- 00:54sgRNA Design
- 01:57Homology-Directed Repair Vector Construction
- 05:42Editing of Hematopoietic Stem and Progenitor Cells
- 09:48Results: Heterozygous Gain-of-Function Mutations in Hematopoietic Stem and Progenitor Cells by Combining the Use of CRISPR/Cas9 and Dual rAAV Donor Transduction
- 11:19Conclusion
Novel strategies to faithfully model somatic mutations in hematopoietic stem and progenitor cells (HSPCs) are necessary to better study hematopoietic stem cell biology and hematological malignancies. Here, a protocol to model heterozygous gain-of-function mutations in HSPCs by combining the use of CRISPR/Cas9 and dual rAAV donor transduction is described.
