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.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Jacobs EZ,Warrier S,Volders PJ,D'haene E,Van Lombergen E,Vantomme L,Van der Jeught M,Heindryckx B,Menten B,Vergult Sdoi
10.1038/s41598-017-16932-ysubject
Has Abstractpub_date
2017-11-30 00:00:00pages
16650issue
1issn
2045-2322pii
10.1038/s41598-017-16932-yjournal_volume
7pub_type
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