RT Journal Article
SR Electronic
T1 Targeted gene correction of <em>FKRP</em> by CRISPR/Cas9 restores functional glycosylation of α-dystroglycan in cortical neurons derived from human induced pluripotent stem cells
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 101352
DO 10.1101/101352
A1 Beatrice Lana
A1 Jihee Kim
A1 David Ryan
A1 Evangelos Konstantinidis
A1 Sandra Louzada
A1 Beiyuan Fu
A1 Fengtang Yang
A1 Derek L. Stemple
A1 Pentao Liu
A1 Francesco Muntoni
A1 Yung-Yao Lin
YR 2017
UL http://biorxiv.org/content/early/2017/01/18/101352.abstract
AB Mutations in genes required for functional glycosylation of α-dystroglycan cause a group of congenital muscular dystrophies associated with brain malformations, referred to as dystroglycanopathies. The lack of isogenic, physiology-relevant human cellular models has limited our understanding of the cortical abnormalities in dystroglycanopathies. Here we generate induced pluripotent stem cells (iPSCs) from a severe dystroglycanopathy patient with homozygous mutations in the ribitol-5-phosphate transferase gene, FKRP. We carry out targeted gene correction in FKRP-iPSCs using CRISPR/Cas9-mediated genome editing. We characterise the directed differentiation of FKRP- and corrected-iPSCs to neural stem cells, cortical progenitors and cortical neurons. Importantly, we show that targeted gene correction of FKRP restores functional glycosylation of α-dystroglycan in iPSC-derived cortical neurons. We independently validate this result by showing targeted gene mutation of FKRP disrupts functional glycosylation of α-dystroglycan. This work demonstrates the feasibility of using CRISPR/Cas9-engineered human iPSCs for modelling dystroglycanopathies and provides a foundation for therapeutic development.HighlightsGeneration of FKRP-iPSCs for modelling cortical abnormalities in dystroglycanopathiesPrecise gene correction by CRISPR/Cas9-mediated genome editingDirected differentiation of isogenic control and FKRP-iPSC to cortical neuronsFunctional glycosylation of α-dystroglycan is restored in cortical neurons derived from CRISPR/Cas9-corrected iPSCsTargeted gene mutation of FKRP disrupts functional glycosylation of α-dystroglycan in cortical neurons