CRISPR/Cas9-mediated genome editing in naïve human embryonic stem cells
| Autor: | Lies Vantomme, Eva D’haene, Margot Van der Jeught, Sarah Vergult, Eva Van Lombergen, Björn Menten, Pieter-Jan Volders, Eva Z. Jacobs, Sharat Warrier, Björn Heindryckx |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Cellular differentiation Human Embryonic Stem Cells lcsh:Medicine Biology DISEASE Article EPIBLAST Cell Line Workflow 03 medical and health sciences Genome editing Neural Stem Cells CRISPR Humans Clustered Regularly Interspaced Short Palindromic Repeats lcsh:Science Genetics Gene Editing HUMAN PLURIPOTENCY Multidisciplinary Cas9 lcsh:R Biology and Life Sciences High-Throughput Nucleotide Sequencing Cell Differentiation LINEAGE Embryonic stem cell STATE Neural stem cell Cell biology DIFFERENTIATION SEQ DERIVATION 030104 developmental biology Epiblast lcsh:Q Stem cell CRISPR-Cas Systems CRISPR CAS9 GENERATION |
| Zdroj: | Scientific Reports SCIENTIFIC REPORTS Scientific Reports, Vol 7, Iss 1, Pp 1-12 (2017) |
| ISSN: | 2045-2322 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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