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| Status |
Public on May 10, 2024 |
| Title |
Proteome-scale tagging and functional screening in mammalian cells by ORFtag [RNA-seq] |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
The systematic determination of protein function is a key goal of modern biology, but remains challenging with current approaches. Here, we present ORFtag, a versatile, cost-effective and highly efficient method for the massively parallel tagging and functional interrogation of proteins at proteome scale. ORFtag utilizes retroviral vectors bearing a promoter, peptide tag and splice donor to generate fusions between the tag and endogenous ORFs. We demonstrate the utility of ORFtag through functional screens for transcriptional activators, repressors and post-transcriptional regulators in mouse embryonic stem cells. Each screen finds known and novel regulators, including long ORFs inaccessible by other methods, revealing that Zfp574 is a highly selective transcriptional activator and that oncogenic fusions often function as transactivators.
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| Overall design |
RNA-Seq of ORFtag activator screen
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| Contributor(s) |
Nemčko F, Himmelsbach M, Loubiere V, Yelagandula R, Pagani M, Fasching N, Brennecke J, Elling U, Stark A, Ameres SL |
| Citation(s) |
38969721 |
| |
| Submission date |
May 09, 2024 |
| Last update date |
Aug 10, 2024 |
| Contact name |
Alexander Stark |
| E-mail(s) |
stark@starklab.org
|
| Organization name |
The Research Institute of Molecular Pathology (IMP)
|
| Lab |
Stark Lab
|
| Street address |
Campus-Vienna-Biocenter 1
|
| City |
Vienna |
| ZIP/Postal code |
1030 |
| Country |
Austria |
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| Platforms (1) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
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| Samples (2) |
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| This SubSeries is part of SuperSeries: |
| GSE225972 |
Proteome-scale tagging and functional screening in mammalian cells by ORFtag |
|
| Relations |
| BioProject |
PRJNA1109780 |
