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CRISPR/Cas9-mediated genome editing in naïve human embryonic stem cells.

CRISPR/Cas9-mediated genome editing in naïve human embryonic stem cells.
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Jacobs EZ, Warrier S, Volders PJ, D'haene E, Van Lombergen E, Vantomme L, Van der Jeught M, Heindryckx B, Menten B, Vergult S,


Jacobs EZ, Warrier S, Volders PJ, D'haene E, Van Lombergen E, Vantomme L, Van der Jeught M, Heindryckx B, Menten B, Vergult S, (click to view)

Jacobs EZ, Warrier S, Volders PJ, D'haene E, Van Lombergen E, Vantomme L, Van der Jeught M, Heindryckx B, Menten B, Vergult S,

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Scientific reports 2017 11 307(1) 16650 doi 10.1038/s41598-017-16932-y
Abstract

The combination of genome-edited human embryonic stem cells (hESCs) and subsequent neural differentiation is a powerful tool to study neurodevelopmental disorders. Since the naïve state of pluripotency has favourable characteristics for efficient genome-editing, we optimized a workflow for the CRISPR/Cas9 system in these naïve stem cells. Editing efficiencies of respectively 1.3-8.4% and 3.8-19% were generated with the Cas9 nuclease and the D10A Cas9 nickase mutant. Next to this, wildtype and genome-edited naïve hESCs were successfully differentiated to neural progenitor cells. As a proof-of-principle of our workflow, two monoclonal genome-edited naïve hESCs colonies were obtained for TUNA, a long non-coding RNA involved in pluripotency and neural differentiation. In these genome-edited hESCs, an effect was seen on expression of TUNA, although not on neural differentiation potential. In conclusion, we optimized a genome-editing workflow in naïve hESCs that can be used to study candidate genes involved in neural differentiation and/or functioning.

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