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

Eva Jacobs (UGent) , Sharat Warrier (UGent) , Pieter-Jan Volders (UGent) , Eva D'haene (UGent) , Eva Van Lombergen, Lies Vantomme (UGent) , Margot Van der Jeught (UGent) , Björn Heindryckx (UGent) , Björn Menten (UGent) and Sarah Vergult (UGent)
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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 naive state of pluripotency has favourable characteristics for efficient genome-editing, we optimized a workflow for the CRISPR/Cas9 system in these naive stem cells. Editing efficiencies of respectively 1.3-8.4% and 3.819% were generated with the Cas9 nuclease and the D10A Cas9 nickase mutant. Next to this, wildtype and genome-edited naive hESCs were successfully differentiated to neural progenitor cells. As a proofof- principle of our workflow, two monoclonal genome-edited naive 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 naive hESCs that can be used to study candidate genes involved in neural differentiation and/or functioning.
Keywords
HUMAN PLURIPOTENCY, CRISPR CAS9, DIFFERENTIATION, LINEAGE, DISEASE, STATE, EPIBLAST, SEQ, GENERATION, DERIVATION

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Chicago
Jacobs, Eva, Sharat Warrier, Pieter-Jan Volders, Eva D’haene, Eva Van Lombergen, Lies Vantomme, Margot Van der Jeught, Björn Heindryckx, Björn Menten, and Sarah Vergult. 2017. “CRISPR/Cas9-mediated Genome Editing in Naïve Human Embryonic Stem Cells.” Scientific Reports 7.
APA
Jacobs, E., Warrier, S., Volders, P.-J., D’haene, E., Van Lombergen, E., Vantomme, L., Van der Jeught, M., et al. (2017). CRISPR/Cas9-mediated genome editing in naïve human embryonic stem cells. SCIENTIFIC REPORTS, 7.
Vancouver
1.
Jacobs E, Warrier S, Volders P-J, D’haene E, Van Lombergen E, Vantomme L, et al. CRISPR/Cas9-mediated genome editing in naïve human embryonic stem cells. SCIENTIFIC REPORTS. 2017;7.
MLA
Jacobs, Eva, Sharat Warrier, Pieter-Jan Volders, et al. “CRISPR/Cas9-mediated Genome Editing in Naïve Human Embryonic Stem Cells.” SCIENTIFIC REPORTS 7 (2017): n. pag. Print.
@article{8541691,
  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 naive state of pluripotency has favourable characteristics for efficient genome-editing, we optimized a workflow for the CRISPR/Cas9 system in these naive stem cells. Editing efficiencies of respectively 1.3-8.4\% and 3.819\% were generated with the Cas9 nuclease and the D10A Cas9 nickase mutant. Next to this, wildtype and genome-edited naive hESCs were successfully differentiated to neural progenitor cells. As a proofof- principle of our workflow, two monoclonal genome-edited naive 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 naive hESCs that can be used to study candidate genes involved in neural differentiation and/or functioning.},
  articleno    = {16650},
  author       = {Jacobs, Eva and Warrier, Sharat and Volders, Pieter-Jan and D'haene, Eva and Van Lombergen, Eva and Vantomme, Lies and Van der Jeught, Margot and Heindryckx, Bj{\"o}rn and Menten, Bj{\"o}rn and Vergult, Sarah},
  issn         = {2045-2322},
  journal      = {SCIENTIFIC REPORTS},
  keyword      = {HUMAN PLURIPOTENCY,CRISPR CAS9,DIFFERENTIATION,LINEAGE,DISEASE,STATE,EPIBLAST,SEQ,GENERATION,DERIVATION},
  language     = {eng},
  pages        = {12},
  title        = {CRISPR/Cas9-mediated genome editing in na{\"i}ve human embryonic stem cells},
  url          = {http://dx.doi.org/10.1038/s41598-017-16932-y},
  volume       = {7},
  year         = {2017},
}

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