PXD001301 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | CRISPR-Cas9-based knockout of the prion protein and its effect on the proteome |
Description | The molecular function of the cellular prion protein (PrPC) and the mechanism by which it may contribute to neurotoxicity in prion diseases and Alzheimer’s disease (AD) are only partially understood. Mouse neuroblastoma Neuro2a cells and, more recently, C2C12 myocytes and myotubes have emerged as popular models for investigating the cellular biology of PrP. Mouse epithelial NMuMG cells might become attractive models for studying the possible involvement of PrP in a morphogenetic program underlying epithelial-to-mesenchymal transitions. Here we describe the generation of PrP knockout clones from these cell lines using CRISPR-Cas9 knockout technology. More specifically, knockout clones were generated with two separate gRNAs targeting recognition sites on opposite strands within the first hundred nucleotides of the Prnp coding sequence. Several PrP knockout clones were isolated and genomic insertions and deletions near the CRISPR-target sites were characterized. Subsequently, deep quantitative global proteome analyses that recorded the relative abundance of > 3000 proteins were undertaken to begin to characterize the molecular consequences of PrP deficiency. The levels of ~120 proteins were shown to reproducibly correlate with the presence or absence of PrP, with most of these proteins as belonging to extracellular components, cell junctions or the cytoskeleton. |
HostingRepository | PRIDE |
AnnounceDate | 2014-11-19 |
AnnouncementXML | Submission_2015-04-01_05:53:10.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Gerold Schmitt-Ulms |
SpeciesList | scientific name: Mus musculus (Mouse); NCBI TaxID: 10090; |
ModificationList | S-pyridylethyl-L-cysteine; monohydroxylated residue; carbamoylated residue |
Instrument | Orbitrap Fusion |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2014-09-10 01:33:17 | ID requested | |
1 | 2014-11-19 00:41:59 | announced | |
⏵ 2 | 2015-04-01 05:53:11 | announced | Updated publication reference for PubMed record(s): 25490046. |
Publication List
Mehrabian M, Brethour D, MacIsaac S, Kim JK, Gunawardana CG, Wang H, Schmitt-Ulms G, CRISPR-Cas9-based knockout of the prion protein and its effect on the proteome. PLoS One, 9(12):e114594(2014) [pubmed] |
Keyword List
curator keyword: Biological |
submitter keyword: prion, CRISPR-Cas9, knockout, kockdown |
Contact List
Gerold Schmitt-Ulms |
contact affiliation | University of Toronto |
contact email | g.schmittulms@utoronto.ca |
lab head | |
Gerold Schmitt-Ulms |
contact affiliation | University of Toronto |
contact email | g.schmittulms@utoronto.ca |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2014/11/PXD001301 |
PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD001301
- Label: PRIDE project
- Name: CRISPR-Cas9-based knockout of the prion protein and its effect on the proteome