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| Status |
Public on Nov 24, 2023 |
| Title |
Analysis of alternative exon-dependent interactome remodelling reveals multitasking functions of gene regulatory factors (Chtop RNA-Seq) |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Alternative splicing is fundamental for the expansion of biological complexity, particularly in the vertebrate nervous system. Increasing evidence indicates that developmental stage and tissue-dependent alternative exons often control protein-protein interactions, yet only a minor fraction of these events has been characterized at the protein level. Using affinity purification-mass spectrometry (AP-MS) we show that approximately 60% of analyzed neural-differential exons in proteins previously implicated in transcriptional regulation result in the gain or loss of interaction partners. Focusing on Chtop and Sap30bp, the neural exons in these proteins unexpectedly remodel interactions with partners associated with mRNA processing and trafficking. Sap30bp additionally controls RNA localization by regulating the splicing of <100 nt ‘mini-introns’ that contribute to the nuclear retention of transcripts. These activities of Chtop and Sap30bp are linked to programs of transcriptomic regulation associated with development of the nervous system. AP-MS is thus a powerful approach for elucidating multifaceted functions of proteins imparted by context-dependent alternative exons.
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| Overall design |
This dataset consists of 35 RNA-seq files associated with the manuscript "Analysis of alternative exon-dependent interactome remodelling reveals multitasking functions of gene regulatory factors" characterizing the effect of Chtop siRNA knockdown and neural/non-neural isoform rescue on transcript nuclear/cytoplasmic localization in Neuro2a cell lines. For each isoform (with or without exon 5), a total of 18 samples were analyzed including non-targeting control (NTC), Chtop siRNA knockdown, and Chtop siRNA knockdown with Doxycycline-induced rescue (using an siRNA-resistant Chtop cDNA) across total/nuclear/cytoplasmic fractions in two biological replicates.
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| Contributor(s) |
Roth JF, Braunschweig U, Lin Z, Larsen B, Weatheritt RJ, Gingras A, Blencowe BJ |
| Citation(s) |
38065061 |
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| Submission date |
Feb 09, 2023 |
| Last update date |
Feb 23, 2024 |
| Contact name |
Ulrich Braunschweig |
| Organization name |
University of Toronto
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| Department |
Donnelly Centre
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| Lab |
Benjamin J. Blencowe
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| Street address |
160 College Street
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| City |
Toronto |
| State/province |
Ontario |
| ZIP/Postal code |
M5S 3E1 |
| Country |
Canada |
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| Platforms (1) |
| GPL24247 |
Illumina NovaSeq 6000 (Mus musculus) |
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| Samples (35)
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| GSM7036356 |
Total_Chtop_mEx5_NTC_BR4: N2a Flp-In rtTA (Chtop_w/oExon5), NT-control, Total frac, Bio rep 1 |
| GSM7036357 |
Total_Chtop_mEx5_NTC_BR6: N2a Flp-In rtTA (Chtop_w/oExon5), NT-control, Total frac, Bio rep 2 |
| GSM7036358 |
Total_Chtop_mEx5_KD_BR4: N2a Flp-In rtTA (Chtop_w/oExon5), siChtop, Total frac, Bio rep 1 |
| GSM7036359 |
Total_Chtop_mEx5_KD_BR6: N2a Flp-In rtTA (Chtop_w/oExon5), siChtop, Total frac, Bio rep 2 |
| GSM7036360 |
Total_Chtop_mEx5_RESC_BR4: N2a Flp-In rtTA (Chtop_w/oExon5), siChtop, +Dox (Chtop mEx5 isof resc), Total frac, Bio rep 1 |
| GSM7036361 |
Total_Chtop_mEx5_RESC_BR6: N2a Flp-In rtTA (Chtop_w/oExon5), siChtop, +Dox (Chtop mEx5 isof resc), Total frac, Bio rep 2 |
| GSM7036362 |
Total_Chtop_pEx5_NTC_BR4: N2a Flp-In rtTA (Chtop_w/Exon5), NT-control, Total frac, Bio rep 1 |
| GSM7036363 |
Total_Chtop_pEx5_NTC_BR6: N2a Flp-In rtTA (Chtop_w/Exon5), NT-control, Total frac, Bio rep 2 |
| GSM7036364 |
Total_Chtop_pEx5_KD_BR4: N2a Flp-In rtTA (Chtop_w/Exon5), siChtop, Total frac, Bio rep 1 |
| GSM7036365 |
Total_Chtop_pEx5_KD_BR6: N2a Flp-In rtTA (Chtop_w/Exon5), siChtop, Total frac, Bio rep 2 |
| GSM7036366 |
Total_Chtop_pEx5_RESC_BR4: N2a Flp-In rtTA (Chtop_w/Exon5), siChtop, +Dox (Chtop pEx5 isof resc), Total frac, Bio rep 1 |
| GSM7036367 |
Total_Chtop_pEx5_RESC_BR6: N2a Flp-In rtTA (Chtop_w/Exon5), siChtop, +Dox (Chtop pEx5 isof resc), Total frac, Bio rep 2 |
| GSM7036368 |
Nuc_Chtop_mEx5_NTC_BR4: N2a Flp-In rtTA (Chtop_w/oExon5), NT-control, nuc frac, Bio rep 1 |
| GSM7036369 |
Nuc_Chtop_mEx5_NTC_BR6: N2a Flp-In rtTA (Chtop_w/oExon5), NT-control, nuc frac, Bio rep 2 |
| GSM7036370 |
Nuc_Chtop_mEx5_KD_BR4: N2a Flp-In rtTA (Chtop_w/oExon5), siChtop, nuc frac, Bio rep 1 |
| GSM7036371 |
Nuc_Chtop_mEx5_KD_BR6: N2a Flp-In rtTA (Chtop_w/oExon5), siChtop, nuc frac, Bio rep 2 |
| GSM7036372 |
Nuc_Chtop_mEx5_RESC_BR4: N2a Flp-In rtTA (Chtop_w/oExon5), siChtop, +Dox (Chtop mEx5 isof resc), nuc frac, Bio rep 1 |
| GSM7036373 |
Nuc_Chtop_mEx5_RESC_BR6: N2a Flp-In rtTA (Chtop_w/oExon5), siChtop, +Dox (Chtop mEx5 isof resc), nuc frac, Bio rep 2 |
| GSM7036374 |
Nuc_Chtop_pEx5_NTC_BR4: N2a Flp-In rtTA (Chtop_w/Exon5), NT-control, nuc frac, Bio rep 1 |
| GSM7036375 |
Nuc_Chtop_pEx5_NTC_BR6: N2a Flp-In rtTA (Chtop_w/Exon5), NT-control, nuc frac, Bio rep 2 |
| GSM7036376 |
Nuc_Chtop_pEx5_KD_BR4: N2a Flp-In rtTA (Chtop_w/Exon5), siChtop, nuc frac, Bio rep 1 |
| GSM7036377 |
Nuc_Chtop_pEx5_KD_BR6: N2a Flp-In rtTA (Chtop_w/Exon5), siChtop, nuc frac, Bio rep 2 |
| GSM7036378 |
Nuc_Chtop_pEx5_RESC_BR4: N2a Flp-In rtTA (Chtop_w/Exon5), siChtop, +Dox (Chtop pEx5 isof resc), nuc frac, Bio rep 1 |
| GSM7036379 |
Nuc_Chtop_pEx5_RESC_BR6: N2a Flp-In rtTA (Chtop_w/Exon5), siChtop, +Dox (Chtop pEx5 isof resc), nuc frac, Bio rep 2 |
| GSM7036380 |
Cyto_Chtop_mEx5_NTC_BR4: N2a Flp-In rtTA (Chtop_w/oExon5), NT-control, cytop frac, Bio rep 1 |
| GSM7036381 |
Cyto_Chtop_mEx5_NTC_BR6: N2a Flp-In rtTA (Chtop_w/oExon5), NT-control, cytop frac, Bio rep 2 |
| GSM7036382 |
Cyto_Chtop_mEx5_KD_BR4: N2a Flp-In rtTA (Chtop_w/oExon5), siChtop, cytop frac, Bio rep 1 |
| GSM7036383 |
Cyto_Chtop_mEx5_KD_BR6: N2a Flp-In rtTA (Chtop_w/oExon5), siChtop, cytop frac, Bio rep 2 |
| GSM7036384 |
Cyto_Chtop_mEx5_RESC_BR4: N2a Flp-In rtTA (Chtop_w/oExon5), siChtop, +Dox (Chtop mEx5 isof resc), cytop frac, Bio rep 1 |
| GSM7036385 |
Cyto_Chtop_mEx5_RESC_BR6: N2a Flp-In rtTA (Chtop_w/oExon5), siChtop, +Dox (Chtop mEx5 isof resc), cytop frac, Bio rep 2 |
| GSM7036386 |
Cyto_Chtop_pEx5_NTC_BR4: N2a Flp-In rtTA (Chtop_w/Exon5), NT-control, cytop frac, Bio rep 1 |
| GSM7036387 |
Cyto_Chtop_pEx5_KD_BR4: N2a Flp-In rtTA (Chtop_w/Exon5), siChtop, cytop frac, Bio rep 1 |
| GSM7036388 |
Cyto_Chtop_pEx5_KD_BR6: N2a Flp-In rtTA (Chtop_w/Exon5), siChtop, cytop frac, Bio rep 2 |
| GSM7036389 |
Cyto_Chtop_pEx5_RESC_BR4: N2a Flp-In rtTA (Chtop_w/Exon5), siChtop, +Dox (Chtop pEx5 isof resc), cytop frac, Bio rep 1 |
| GSM7036390 |
Cyto_Chtop_pEx5_RESC_BR6: N2a Flp-In rtTA (Chtop_w/Exon5), siChtop, +Dox (Chtop pEx5 isof resc), cytop frac, Bio rep 2 |
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| This SubSeries is part of SuperSeries: |
| GSE224971 |
Analysis of alternative exon-dependent interactome remodelling reveals multitasking functions of gene regulatory factors |
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| Relations |
| BioProject |
PRJNA934536 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE224969_NCFracts_Chtop_COUNTS.txt.gz |
685.2 Kb |
(ftp)(http) |
TXT |
| GSE224969_NCFracts_Chtop_DESeq2_normCOUNTS_CytovsTotal_wAnnotations.txt.gz |
2.5 Mb |
(ftp)(http) |
TXT |
| GSE224969_NCFracts_Chtop_DESeq2_normCOUNTS_NucvsCyto_wAnnotations.txt.gz |
2.5 Mb |
(ftp)(http) |
TXT |
| GSE224969_NCFracts_Chtop_DESeq2_normCOUNTS_NucvsTotal_wAnnotations.txt.gz |
2.5 Mb |
(ftp)(http) |
TXT |
| GSE224969_NCFracts_Chtop_RPKM.tab.gz |
1.9 Mb |
(ftp)(http) |
TAB |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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