CRISPR/Cas9-mediated Genome Editing in Naïve Human Embryonic Stem Cells

Overview

Journal Sci Rep

Specialty Science

Date 2017 Dec 2

PMID 29192200

Citations 4

Authors

Eva Z Jacobs

Sharat Warrier

Pieter-Jan Volders

Eva Dhaene

Eva Van Lombergen

Lies Vantomme

Margot van der Jeught

Bjorn Heindryckx

Bjorn Menten

Sarah Vergult

Affiliations

Soon will be listed here.

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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