Summary: Introduction To Stemcells

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  • 1 Day 1: Embryonic stem cells, pluripotency and reprogramming

  • 1.1 From embryo to embryonic stem cells (Roelen)

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  • Oocyte (egg cell)

    Large immotile cell.
  • Which tissues can a pluripotent stem cells differentiate towards?

    Endoderm, mesoderm, germ cells (NO yolk sack and placenta).
  • 1.3 Reprogramming, induced pluripotent stem cells and application (Jaenisch)

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  • Sporadic, late onset diseases

    • Common and therefore medically the most relevant
    • Lack a clear genetic basis
    • Generally display a more subtle phenotype   
  • Therapeutic applications of iPSCs?

    • Can be used to generate desired cell types for individual patients
    • Can be used to model disease, facilitating drug discovery.
    • Patient-derived iPSCs could potentially be corrected ex vivo to be transplanted back into the patient to replenish lost/damaged cells.   
  • Major issue with iPSC-derived cells

    They often remain poorly matured
  • How can we manipulate the human (epi)genome to rectify disease-causing variants?

    • Gene targeting
    • CRISPR/Cas9-bases gene editing
  • CRISPR/Cas9-based gene editing

    • Efficient method to generate/correct mutations, based on HR on nonhomologous end joining (NHEJ)
    • Involves a guide RNA (for homology search) and Cas9 (cleaves DNA).
    • Can introduce multiple genetic modifications at a time.
    • Efficient and rapid technique.
    • Does not require extensive selection process.    
  • Two main types of gene therapies?

    • Somatic therapy
    • Germline therapy
  • 2 Day 2: Epigenetics and flavors of pluripotency

  • 2.1 Principles of epigenetics in stem cells (Lopes)

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  • Association of ATP-dependent complexes

    • E.g. polycomb and trithorax proteins
    • Change the distance between nucleosomes, which results in altered accessibility 
  • Association with non-coding RNAs

    • Binding of non-coding RNAs to the DNA blocks transcription
    • E.g. X-inactive specific transcript (XIST) and TSIX
      •  XIST and TSIX can neutralize each other 

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