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| Status |
Public on Feb 24, 2021 |
| Title |
Gene expression profile of iPSC-derived dopaminergic neurons from control and Parkinson's disease patients carrying LRRK2 G2019S point mutation |
| Organism |
Homo sapiens |
| Experiment type |
Other
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| Summary |
The goal of this study is to compare the RNA expression profile from human iPSC-derived dopaminergic neurons from control, Parkinson’s disease LRRK2 G2019S and gene-edited isogenic lines (with corrected LRRK2 mutation) to gain insight into the mechanisms underlying neuronal degeneration in PD.
Total RNA was assayed for quantity and quality using Qubit® RNA HS Assay (Life Technologies) and RNA 6000 Nano Assay on a Bioanalyzer 2100. All the steps were carried out in the ice and using the nCounter nanoString reagent for the Neuropathology panel. The master mix for the hybridization process was prepared by mixing hybridization buffer and reporter CodeSet. A dilution of 10μg/μl for each RNA to be analyzed was prepared and maintained in ice. For each sample, a tube was prepared with 5μl of RNA (10μg/μl) and 8μl of master mix. 2μl of Capture ProbeSet was added in each tube and the hybridization process was started using a PCR machine at 65ºC for at least 16 hours. After this step, the samples were maintained at 4ºC. Once the hybridization process was completed, the hybridized RNA was loaded in the neuropathology cartridge, and the data were available 24 hours later.
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| Overall design |
Parkinson's disease (PD) human iPSC-derived neuronal expression profile was obtained by comparing human iPSC-derived neurons from control, PD LRRK2 G2019S patients and isogenic lines. MRNA profiles were generated using the nCounter nanoString reagent for Neuropathology panel. Control = from one healthy individual; 2 independent differentiations. PD = from 2 PD-LRRK2 G2019S patients; 2/3 independent differentiations; isoPD = from 2 corrected PD-LRRK2 patients; 2/3 independent differentations.
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| Contributor(s) |
Pons M |
| Citation(s) |
34215735 |
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| Submission date |
Feb 23, 2021 |
| Last update date |
Jul 27, 2021 |
| Contact name |
Meritxell Pons |
| E-mail(s) |
meritxell.pons@ub.edu
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| Organization name |
IDIBELL
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| Lab |
Stem Cells and Neurodegeneration Lab
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| Street address |
Avinguda Gran Via, 199-203
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| City |
Hospitalet de Llobregat (Barcelona) |
| ZIP/Postal code |
08908 |
| Country |
Spain |
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| Platforms (1) |
| GPL29761 |
Nanostring nCounter Human Neuropathology Panel |
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| Samples (10)
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| Relations |
| BioProject |
PRJNA704331 |
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