PXD012613 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | TRIM21-mediated protein depletion recapitulates mutant gene phenotype in early mouse embryos |
Description | It is often desirable to be able to regulate or impair the expression of specific genes to study early developmental events in the mammalian embryo. While DNA and RNA methods are routine, methods using proteins are still in their infancy. When proteins in the cell encounter a specific antibody and the ubiquitin-protein ligase TRIM21, a ternary complex forms with the target protein, leading to its rapid and acute degradation – hence the name ‘Trim-Away'. However, there are many unknowns in this new endeavour. First and foremost, the extent to which endogenous proteins can be depleted depends on their amount in relation to the amount of exogenous antibody, which is limited by the microinjection procedure. Secondly, the depletion of the protein must be sustained over days. Using mass spectrometry and the iBAQ algorithm, we estimate the amount of proteins found in preimplantation mouse embryos. Most of these amounts are tractable with the microinjection method presented here, which supplies 10E-4 picomoles of antibody contained in 100 picolitres, before incurring toxic effects on mouse development. Building on these data, we demonstrate the feasibility of protein knock-down for a gene which is essential in the preimplantation mouse embryo, namely TEA domain family member 4 (Tead4). Knock-down persists long enough to result in a phenotype which is entirely consistent with that of the null mutation and the RNA interference: significantly reduced mRNA expression of TEAD4 target genes Cdx2 and Gata3, failure of CDX2 nuclear translocation and the embryo’s inability to implant. We conclude that at least for a time window of 3-4 days of preimplantation development, protein depletion is on the rise as a valid alternative to DNA and RNA methods. |
HostingRepository | PRIDE |
AnnounceDate | 2024-10-22 |
AnnouncementXML | Submission_2024-10-22_04:52:54.863.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Hannes Drexler |
SpeciesList | scientific name: Mus musculus (Mouse); NCBI TaxID: 10090; |
ModificationList | monohydroxylated residue; acetylated residue |
Instrument | LTQ Orbitrap Velos; Q Exactive |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2019-02-07 03:54:09 | ID requested | |
1 | 2019-09-11 08:48:03 | announced | |
2 | 2019-10-25 04:12:14 | announced | 2019-10-25: Updated publication reference for PubMed record(s): 31638890. |
⏵ 3 | 2024-10-22 04:53:03 | announced | 2024-10-22: Updated project metadata. |
Publication List
Israel S, Casser E, Drexler HCA, Fuellen G, Boiani M, A framework for TRIM21-mediated protein depletion in early mouse embryos: recapitulation of Tead4 null phenotype over three days. BMC Genomics, 20(1):755(2019) [pubmed] |
10.1186/s12864-019-6106-2; |
Keyword List
curator keyword: Biological |
submitter keyword: mouse, embryo, early development, SILAC, iBAQ, oocyte, proteome |
Contact List
Hannes C. A. Drexler |
contact affiliation | Max Planck Institut for Molecular Biomedicine Bioanalytical Mass Spectrometry Röntgenstr. 20 48159 Münster Germany |
contact email | hannes.drexler@mpi-muenster.mpg.de |
lab head | |
Hannes Drexler |
contact affiliation | Bioanalytical Mass Spectrometry, Max Planck Insitute for Molecular Biomedicine, Roentgenstr. 20, 48149 Münster, Germany |
contact email | hannes.drexler@mpi-muenster.mpg.de |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD012613
- Label: PRIDE project
- Name: TRIM21-mediated protein depletion recapitulates mutant gene phenotype in early mouse embryos