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| Status |
Public on Dec 15, 2019 |
| Title |
Determination of Sox10-regulated genes in the oligodendroglial precursor cell line OLN93 |
| Organism |
Rattus norvegicus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
To determine Sox10-regulated genes in the oligodendroglial precursor cell line OLN93, CRISPR/Cas9-mediated genome editing was performed to inactivate Sox10. For CRISPR/Cas9-mediated Sox10 inactivation, two guide sequences (5’-GCTGCGAGCCAGCCCAGGGCC-3’ for clone A and 5’-GTTGCCCAATTCGCCG GGCCC-3’ for clone B) were separately cloned into pX330. Oln93 cells were co-transfected with pEGFP-N1 and the pX330 plasmids that express the respective guide RNA and SpCas9. Transfected GFP-positive cells were enriched by fluorescence activated cell-sorting and seeded at single cell density in 96-well plates. Homogeneously GFP-expressing clones were analysed for loss of Sox10 protein by immunocytochemistry. Clones without Sox10 protein were obtained with both guide RNAs. Sox10 inactivation was confirmed by Sanger sequencing of the Sox10 gene. RNA was isolated from one clone A and one clone B as well as two independent RNA samples of wildtype OLN93 cells and submitted to RNA-seq.
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| Overall design |
mRNA profiles of control (ctrl) and genome-edited Sox10-deficient (Sox10ko) OLN93 cells were generated using Illumina HiSeq 2500 platform.
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| Contributor(s) |
Weider M, Wegner M, Fröb F |
| Citation(s) |
31828317 |
| Submission date |
Aug 29, 2019 |
| Last update date |
Mar 02, 2020 |
| Contact name |
Michael Wegner |
| Organization name |
Friedrich-Alexander Universität Erlangen-Nürnberg
|
| Department |
Institut of Biochemistry
|
| Lab |
Michael Wegner
|
| Street address |
Fahrstr. 17
|
| City |
Erlangen |
| State/province |
Bavaria |
| ZIP/Postal code |
91054 |
| Country |
Germany |
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| Platforms (1) |
| GPL18694 |
Illumina HiSeq 2500 (Rattus norvegicus) |
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| Samples (4)
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| GSM4054734 |
OLN93 control (ctrl) cells, biological replicate 1 |
| GSM4054735 |
OLN93 control (ctrl) cells, biological replicate 2 |
| GSM4054736 |
OLN93 Sox10-deficient (Sox10ko) cells, clone A |
| GSM4054737 |
OLN93 Sox10-deficient (Sox10ko) cells, clone B |
|
| Relations |
| BioProject |
PRJNA562933 |
| SRA |
SRP219661 |
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