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Optimized Platform Boosts Studies of Rare Neurodevelopmental Disorders

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Review of “A Rapid Pipeline to Model Rare Neurodevelopmental Disorders with Simultaneous CRIPSR/Cas9 Gene Editing” from Stem Cells Translational Medicine by Stuart P. Atkinson

Advances in patient- and disease-specific induced pluripotent stem cell (iPSC) technology promises to greatly advanced our understanding of disease cause, progression, and possible treatment strategies by modeling common neurodevelopmental disorders.

In order to potentiate these advances and extend the technology to rarer neurodevelopmental disorders [1], the lab of Carl Ernst (McGill University, Quebec, Canada) has created an optimized platform for homogenous iPSCs production, accurate CRISPR/Cas9 gene-editing, and efficient neuronal differentiation in a time- and cost-effective manner [2]. 

Will this new strategy, reported in Stem Cells Translational Medicine, lead to an explosion of new models, new findings, and new treatments for rare neurodevelopmental disorders?

The optimized platform developed by the team involved several interrelated steps:

  • Quick and efficient iPSC generation via the transfection of skin fibroblasts with episomal iPSC induction vectors also containing a puromycin resistance gene
    • An initial 24 hour puromycin selection stage enabled rapid reprogramming and generated iPSC colonies on Matrigel-coated plates in mTesR-E7 in 14 to 20 days
    • This efficient process generated 18 to 24 pure colonies per 2,000 cells plated
    • Pure colonies generated by day 25 by gentle dissociation and replating utilizing ReLeSR media (enzyme-free reagent for passaging without manual selection or scraping) 
  • Simultaneous gene editing via the addition of an episomal CRIPSR/CAS9 construct and a specific gRNA alongside the reprogramming vectors
    • Gene editing did not affect reprogramming and easily produced homozygous and heterozygous iPSCs lines
    • Gene editing in fibroblasts represents an easier task than in iPSCs and allows for single cell work, so allowing the production of differentiated cells derived from a single, edited fibroblast, ensuring genotypically homogenous cells
    • Gene editing permits modeling of monogenic diseases when patient tissue is unavailable, testing of genetic contributions to complex diseases, and generation of isogenic control iPSCs 
  • Generation of forebrain and midbrain neural cells via efficient directed differentiation
    • Three-dimensional differentiation of iPSCs as organoids permitted the generation of physiologically active forebrain GABA/glutamatergic neurons and midbrain dopaminergic neurons in around 2 months
    • Strategy also permitted the isolation and frozen storage of neural progenitor cells at specific stages without affecting subsequent differentiation potential

Quick, cheap, simple, and reliable; what’s not to like? This optimized platform combining rapid reprogramming, accurate gene-editing, and efficient neuronal differentiation could prove to be an immense resource for the study of rare neurodevelopmental disorders. Furthermore, modifications to this system may allow the production of cells of other lineages for developmental studies, drug screens, or even direct cell therapies.

Stay tuned to the Stem Cell Portal to see just how far this platform can go!


  1. Bishop DVM. Which Neurodevelopmental Disorders Get Researched and Why? PLOS ONE 2010;5:e15112.
  2. Bell S, Peng H, Crapper L, et al. A Rapid Pipeline to Model Rare Neurodevelopmental Disorders with Simultaneous CRISPR/Cas9 Gene Editing. Stem Cells Transl Med 2017;6:886-896.