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Hemogenic Reprogramming of Human Fibroblasts by Enforced Expression of Transcription Factors
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Increased error-free DNA repair gene expression through reprogramming in human iPS cells.

Yasuhide Yoshimura1

  • 1Division of Gene Therapy Science, Department of Genome Biology, Graduate School of Medicine, Osaka University, Address: 2-2 Yamada-oka, Suita, Osaka 565-0871, Japan.

Regenerative Therapy
|July 16, 2019
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Summary

Human-induced pluripotent stem cells (hiPSCs) enhance DNA repair mechanisms, including homologous recombination, during reprogramming. This boosts precise genome editing but may increase cell death risk.

Keywords:
BLMBLM, the gene defective in Bloom's syndromeDNA repairHDF, human dermal fibroblastHomologous recombinationMSC, mesenchymal stromal cellPARPPARP, poly (ADP-ribose) polymeraseRAD51ROS, Reactive Oxygen SpeciesReprogrammingiPS, induced pluripotent stem

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Area of Science:

  • Stem cell biology
  • DNA repair mechanisms
  • Regenerative medicine

Background:

  • Human-induced pluripotent stem cells (hiPSCs) and human embryonic stem cells (hESCs) exhibit remarkable DNA repair capabilities.
  • Unlike differentiated cells, hESCs possess mechanisms like apoptosis-prone mitochondria to prevent transmission of genetic alterations.
  • While genomic changes during pluripotency are studied, alterations in DNA repair post-reprogramming remain less understood.

Purpose of the Study:

  • To investigate the changes in DNA repair gene expression and activity in hiPSCs during reprogramming.
  • To understand how reprogramming influences the DNA repair pathways utilized by pluripotent stem cells.

Main Methods:

  • Microarray analysis of DNA repair-related genes in hiPSCs.
  • Western blotting to assess protein expression.
  • Measurement of poly (ADP-ribose) polymerase (PARP) activity changes during reprogramming.

Main Results:

  • Reprogramming upregulated PARP activity and homologous recombination (HR) gene expression in hiPSCs.
  • Non-homologous end joining (NHEJ) gene expression remained low, suggesting HR is the preferred repair pathway for DNA scission.
  • Mismatch repair (MMR) genes were upregulated, consistent with high proliferative activity, while error-prone polymerases were downregulated.

Conclusions:

  • Reprogramming induces high PARP activity and HR gene expression in hiPSCs, facilitating precise genome editing.
  • These enhanced DNA repair mechanisms may come at the cost of an increased risk of cell death.