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Engineering Oncogenic Heterozygous Gain-of-Function Mutations in Human Hematopoietic Stem and Progenitor Cells
JoVE 杂志
癌症研究
This content is Free Access.
JoVE 杂志 癌症研究
Engineering Oncogenic Heterozygous Gain-of-Function Mutations in Human Hematopoietic Stem and Progenitor Cells
DOI:

12:04 min

March 10, 2023

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Chapters

  • 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

Summary

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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.

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