In vivo CRISPR/Cas9-mediated screen reveals a critical function of TFDP1 and E2F4 transcription factors in hematopoiesis

Leukemia. 2024 Sep;38(9):2003-2015. doi: 10.1038/s41375-024-02357-w. Epub 2024 Jul 23.

Abstract

Hematopoiesis is a continuous process of blood cell production driven by hematopoietic stem and progenitor cells (HSPCs) in the bone marrow. Proliferation and differentiation of HSPCs are regulated by complex transcriptional networks. In order to identify transcription factors with key roles in HSPC-mediated hematopoietic reconstitution, we developed an efficient and robust CRISPR/Cas9-based in vivo genetic screen. Using this experimental system, we identified the TFDP1 transcription factor to be essential for HSPC proliferation and post-transplant hematopoiesis. We further discovered that E2F4, an E2F transcription factor, serves as a binding partner of TFDP1 and is required for HSPC proliferation. Deletion of TFDP1 caused downregulation of genes associated with the cell cycle, with around 50% of these genes being identified as direct targets of TFDP1 and E2F4. Thus, our study expands the transcriptional network governing hematopoietic development through an in vivo CRISPR/Cas9-based genetic screen and identifies TFDP1/E2F4 as positive regulators of cell cycle genes in HSPCs.

MeSH terms

  • Animals
  • CRISPR-Cas Systems*
  • Cell Cycle / genetics
  • Cell Differentiation / genetics
  • Cell Proliferation
  • E2F4 Transcription Factor* / genetics
  • E2F4 Transcription Factor* / metabolism
  • Hematopoiesis* / genetics
  • Hematopoietic Stem Cells* / cytology
  • Hematopoietic Stem Cells* / metabolism
  • Humans
  • Mice
  • Mice, Inbred C57BL
  • Transcription Factor DP1* / genetics
  • Transcription Factor DP1* / metabolism

Substances

  • E2f4 protein, mouse
  • E2F4 Transcription Factor
  • Transcription Factor DP1
  • Tfdp1 protein, mouse