Functional genomics

Florence van TienenFunctional genomics is a (part novel) activity within Clinical Genetics. Functional genomics uses several tools to characterize nucleotide variants with an unknown effect (VUS). We use known models or new models.

iPSC reprogramming

Pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), are able to self-renew indefinitely and differentiate into a wide spectrum of cell types. For iPSCs generation, various cell-types can be used, but most often fibroblasts are used, since they can be relatively easily obtained.

What do we offer?

  • Generate iPSC clones of fibroblasts (or other cell-types upon request) using lentiviral method
  • Analysis of pluripotency and differentiation capacity of iPSC clones
  • Karyotype analysis of iPSC clones

Pericyte/Mesoangioblasts (MABs) culture from a muscle biopsy

Pericytes, also called mesoangioblasts (MABs), can be isolated form a muscle biopsy and expanded. Mesoangioblasts can proliferate for >20 population doublings and can spontaneously differentiate into myotubes. Mesoangioblasts can also be reprogrammed to iPSC and they can also directly differentiate into other cell-types of the mesoderm, like adipocytes and smooth muscle.

What do we offer?

  • Isolation and characterization of mesoangioblasts from fresh skeletal muscle biopsies
  • Differentiation of mesoangioblasts into myotubes

Direct differentiation of fibroblasts into myotubes

To enable (RNA) analysis of muscle-specific splice variants, (patient)fibroblasts can be directly differentiated into myotubes by MyoD overexpression.

Jo VanoevelenWhat do we offer?

  • Generate fibroblast culture from skin biopsy
  • Generate myotubes from fibroblasts by MyoD overexpression
  • RNA analysis of muscle specific transcripts

Stemcell derived models

  • iPSC or ES differentiation to cardiomyocytes
  • iPSC or ES differentiation to neuronic precursors and differentiation to neuronal cells (location¬†Nijmegen)

Tools (Morpholino-mediated knockdown)

  • Knock-out lines (CrisPR/Cas9)
  • Knock-in lines (CrisPR/Cas9)
  • Reporter lines (Tol2 transgenesis)

Genetic engineering (of cell and animal models)

  • Crisp/Cas
  • DNA cloning, introduction of mutations and plasmid construction

Contact

When planning an experiment involving functional genomics, feel free to contact us for an initial consult.

Florence van Tienen, PhD (iPSC models)
Jo Vanoevelen, PhD (zebrafish models)
E: gsm@mumc.nl