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GSE232597

GSE GEO
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Systematic identification of RNA-binding proteins and tethered domains that activate exon splicing inclusion [eCLIP-seq]

Organism: Homo sapiens
Platform: GPL24676
Samples: 20
Experiment Types:
Other
Submitted: May 16 2023
Last Updated: Oct 08 2024
Status: Public on Sep 25 2023
Contact: Brian,,Yee (UCSD)

Relations

SubSeries of: GSE232599 BioProject: https://www.ncbi.nlm.nih.gov/bioproject/PRJNA972979

Summary

RNA-binding proteins (RBPs) modulate alternative splicing outcomes to determine isoform expression and cellular survival. To identify RBPs that directly drive alternative exon inclusion, we evaluated 718 human RBPs with tethered function luciferase-based splicing reporter assays to identify 58 candidates, including known splicing factors such as RBFOX and serine-arginine proteins. We performed enhanced CLIP, RNA-seq, and affinity purification-mass spectrometry to investigate a subset of the 11 candidates with no prior association with splicing. Integrative analysis of these assays indicated the surprising roles of TRNAU1AP, SCAF8, and RTCA in modulating hundreds of endogenous splicing events. We also leveraged our tethering assays and top candidates to identify potent and compact exon inclusion activation domains for splicing modulation applications. Using identified domains, we engineered programmable fusion proteins which outperformed current artificial splicing factors at manipulating inclusion of reporter and endogenous exons. Altogether, our tethering approach characterized the ability of RBPs to induce exon inclusion and yielded new molecular parts for programmable splicing control.

Overall Design

eCLIP-seq with TRNAU1AP-specific antibody in HEK cells, or V5-specific antibody in HEK cells overexpressing V5-tagged targets

Analysis (5 steps)

View Data Processing
Processing steps for GSE232597
  1. Data was processed using Skipper, available at: http://github.com/yeolab/skipper
  2. Reads were trimmed for adapter sequences and barcode sequences (eCLIP samples) using skewer.
  3. Unique Molecular Identifiers (UMIs) were extracted from raw sequencing reads with fastp
  4. Extracted reads were aligned using STAR: --alignEndsType EndToEnd --genomeDir {params.star_sjdb} --genomeLoad NoSharedMemory --outBAMcompression 10 --outFileNamePrefix {params.outprefix} --winAnchorMultimapNmax 100 --outFilterMultimapNmax 100 --outFilterMultimapScoreRange 1 --outSAMmultNmax 1 --outMultimapperOrder Random --outFilterScoreMin 10 --outFilterType BySJout --limitOutSJcollapsed 5000000 --outReadsUnmapped None --outSAMattrRGline ID:{wildcards.replicate_label} --outSAMattributes All --outSAMmode Full --outSAMtype BAM Unsorted --outSAMunmapped Within --readFilesCommand zcat --outStd Log --readFilesIn {input.fq} --runMode alignReads --runThreadN {threads}
  5. Custom scripts called reproducible enriched windows and repetitive elements as part of Skipper

Supplementary Files (15)

GSE232597_FLAG_IP1_enriched_windows.tsv.gz Download
GSE232597_FLAG_IP2_enriched_windows.tsv.gz Download
GSE232597_FLAG_reproducible_enriched_windows.tsv.gz Download
GSE232597_RTCA_IP1_enriched_windows.tsv.gz Download
GSE232597_RTCA_IP2_enriched_windows.tsv.gz Download
GSE232597_RTCA_reproducible_enriched_windows.tsv.gz Download
GSE232597_SCAF8_IP1_enriched_windows.tsv.gz Download
GSE232597_SCAF8_IP2_enriched_windows.tsv.gz Download
GSE232597_SCAF8_reproducible_enriched_windows.tsv.gz Download
GSE232597_STAU2_IP1_enriched_windows.tsv.gz Download
GSE232597_STAU2_IP2_enriched_windows.tsv.gz Download
GSE232597_STAU2_reproducible_enriched_windows.tsv.gz Download
GSE232597_TRNAU1AP_IP1_enriched_windows.tsv.gz Download
GSE232597_TRNAU1AP_IP2_enriched_windows.tsv.gz Download
GSE232597_TRNAU1AP_reproducible_enriched_windows.tsv.gz Download
GEO Samples (20)

Dataset Citations (1)

Large-scale evaluation of the ability of RNA-binding proteins to activate exon inclusion.
PMID 38168984 · 2024 · Nature biotechnology
Jonathan C Schmok, Manya Jain, Lena A Street, Alex T Tankka, Danielle Schafer, Hsuan-Lin Her, Sara Elmsaouri, Maya L Gosztyla, Evan A Boyle, Pratibha Jagannatha, En-Ching Luo, Ester J Kwon, Marko Jovanovic, Gene W Yeo

SRA Experiments (20) and Runs (20)

Total: 16188 MB
SRX20362686 SRP437920 RIP-Seq SINGLE
GSM7359773: HEK293T cells, TRNAU1AP_IN_1; Homo sapiens; RIP-Seq
Sample: SRS17679614
BioProject: PRJNA972979
BioSample: SAMN35102719
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: No treatment
rip antibody: none
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579347 20833620 2104195620 652.87 TRNAU1AP_IN1.fastq.gz, SRR24579347, SRR24579347.lite SRA
SRX20362687 SRP437920 RIP-Seq SINGLE
GSM7359774: HEK293T cells, TRNAU1AP_IN_2; Homo sapiens; RIP-Seq
Sample: SRS17679615
BioProject: PRJNA972979
BioSample: SAMN35102718
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: No treatment
rip antibody: none
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579346 15672441 1582916541 493.3 TRNAU1AP_IN2.fastq.gz, SRR24579346, SRR24579346.lite SRA
SRX20362688 SRP437920 RIP-Seq SINGLE
GSM7359775: HEK293T cells, TRNAU1AP_IP_1; Homo sapiens; RIP-Seq
Sample: SRS17679616
BioProject: PRJNA972979
BioSample: SAMN35102717
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: No treatment
rip antibody: TRNAU1AP-specific antibody
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579345 16806219 1697428119 514.11 TRNAU1AP_IP1.fastq.gz, SRR24579345, SRR24579345.lite SRA
SRX20362689 SRP437920 RIP-Seq SINGLE
GSM7359776: HEK293T cells, TRNAU1AP_IP_2; Homo sapiens; RIP-Seq
Sample: SRS17679617
BioProject: PRJNA972979
BioSample: SAMN35102716
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: No treatment
rip antibody: TRNAU1AP-specific antibody
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579344 26663497 2693013197 819.35 TRNAU1AP_IP2.fastq.gz, SRR24579344, SRR24579344.lite SRA
SRX20362690 SRP437920 RIP-Seq SINGLE
GSM7359777: HEK293T cells, RTCA_IN_1; Homo sapiens; RIP-Seq
Sample: SRS17679618
BioProject: PRJNA972979
BioSample: SAMN35102715
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: Overexpression of V5-tagged RTCA
rip antibody: none
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579343 38684019 2901301425 824.83 RTCA_IN1.fastq.gz, SRR24579343, SRR24579343.lite SRA
SRX20362691 SRP437920 RIP-Seq SINGLE
GSM7359778: HEK293T cells, RTCA_IN_2; Homo sapiens; RIP-Seq
Sample: SRS17679619
BioProject: PRJNA972979
BioSample: SAMN35102714
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: Overexpression of V5-tagged RTCA
rip antibody: none
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579342 37878702 2840902650 808.68 RTCA_IN2.fastq.gz, SRR24579342, SRR24579342.lite SRA
SRX20362692 SRP437920 RIP-Seq SINGLE
GSM7359779: HEK293T cells, RTCA_IP_1; Homo sapiens; RIP-Seq
Sample: SRS17679620
BioProject: PRJNA972979
BioSample: SAMN35102713
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: Overexpression of V5-tagged RTCA
rip antibody: V5-specific antibody
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579341 38085130 2856384750 816.45 RTCA_IP1.fastq.gz, SRR24579341, SRR24579341.lite SRA
SRX20362693 SRP437920 RIP-Seq SINGLE
GSM7359780: HEK293T cells, RTCA_IP_2; Homo sapiens; RIP-Seq
Sample: SRS17679621
BioProject: PRJNA972979
BioSample: SAMN35102712
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: Overexpression of V5-tagged RTCA
rip antibody: V5-specific antibody
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579340 40049683 3003726225 858.7 RTCA_IP2.fastq.gz, SRR24579340, SRR24579340.lite SRA
SRX20362694 SRP437920 RIP-Seq SINGLE
GSM7359781: HEK293T cells, SCAF8_IN_1; Homo sapiens; RIP-Seq
Sample: SRS17679622
BioProject: PRJNA972979
BioSample: SAMN35102711
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: Overexpression of V5-tagged SCAF8
rip antibody: none
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579339 35063895 2629792125 746.9 SCAF8_IN1.fastq.gz, SRR24579339 SRA
SRX20362695 SRP437920 RIP-Seq SINGLE
GSM7359782: HEK293T cells, SCAF8_IN_2; Homo sapiens; RIP-Seq
Sample: SRS17679623
BioProject: PRJNA972979
BioSample: SAMN35102710
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: Overexpression of V5-tagged SCAF8
rip antibody: none
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579338 41407652 3105573900 882.84 SCAF8_IN2.fastq.gz, SRR24579338, SRR24579338.lite SRA
SRX20362696 SRP437920 RIP-Seq SINGLE
GSM7359783: HEK293T cells, SCAF8_IP_1; Homo sapiens; RIP-Seq
Sample: SRS17679624
BioProject: PRJNA972979
BioSample: SAMN35102709
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: Overexpression of V5-tagged SCAF8
rip antibody: V5-specific antibody
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579337 36476437 2735732775 782.86 SCAF8_IP1.fastq.gz, SRR24579337, SRR24579337.lite SRA
SRX20362697 SRP437920 RIP-Seq SINGLE
GSM7359784: HEK293T cells, SCAF8_IP_2; Homo sapiens; RIP-Seq
Sample: SRS17679625
BioProject: PRJNA972979
BioSample: SAMN35102708
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: Overexpression of V5-tagged SCAF8
rip antibody: V5-specific antibody
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579336 39933385 2995003875 852.63 SCAF8_IP2.fastq.gz, SRR24579336, SRR24579336.lite SRA
SRX20362698 SRP437920 RIP-Seq SINGLE
GSM7359785: HEK293T cells, STAU2_IN_1; Homo sapiens; RIP-Seq
Sample: SRS17679626
BioProject: PRJNA972979
BioSample: SAMN35102707
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: Overexpression of V5-tagged STAU2
rip antibody: none
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579335 37690833 2826812475 805.71 STAU2_IN1.fastq.gz, SRR24579335, SRR24579335.lite SRA
SRX20362699 SRP437920 RIP-Seq SINGLE
GSM7359786: HEK293T cells, STAU2_IN_2; Homo sapiens; RIP-Seq
Sample: SRS17679627
BioProject: PRJNA972979
BioSample: SAMN35102706
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: Overexpression of V5-tagged STAU2
rip antibody: none
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579334 42305202 3172890150 902.56 STAU2_IN2.fastq.gz, SRR24579334, SRR24579334.lite SRA
SRX20362700 SRP437920 RIP-Seq SINGLE
GSM7359787: HEK293T cells, STAU2_IP_1; Homo sapiens; RIP-Seq
Sample: SRS17679628
BioProject: PRJNA972979
BioSample: SAMN35102705
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: Overexpression of V5-tagged STAU2
rip antibody: V5-specific antibody
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579333 43171073 3237830475 925.04 STAU2_IP1.fastq.gz, SRR24579333, SRR24579333.lite SRA
SRX20362701 SRP437920 RIP-Seq SINGLE
GSM7359788: HEK293T cells, STAU2_IP_2; Homo sapiens; RIP-Seq
Sample: SRS17679629
BioProject: PRJNA972979
BioSample: SAMN35102704
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: Overexpression of V5-tagged STAU2
rip antibody: V5-specific antibody
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579332 40871112 3065333400 876.95 STAU2_IP2.fastq.gz, SRR24579332, SRR24579332.lite SRA
SRX20362706 SRP437920 RIP-Seq SINGLE
GSM7359793: HEK293T cells, FLAG_IN_1; Homo sapiens; RIP-Seq
Sample: SRS17679634
BioProject: PRJNA972979
BioSample: SAMN35102699
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: Overexpression of V5-tagged FLAG
rip antibody: none
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579327 43147058 3236029350 928.86 FLAG_IN1.fastq.gz, SRR24579327, SRR24579327.lite SRA
SRX20362707 SRP437920 RIP-Seq SINGLE
GSM7359794: HEK293T cells, FLAG_IN_2; Homo sapiens; RIP-Seq
Sample: SRS17679635
BioProject: PRJNA972979
BioSample: SAMN35102698
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: Overexpression of V5-tagged FLAG
rip antibody: none
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579326 47732648 3579948600 1028.85 FLAG_IN2.fastq.gz, SRR24579326, SRR24579326.lite SRA
SRX20362708 SRP437920 RIP-Seq SINGLE
GSM7359795: HEK293T cells, FLAG_IP_1; Homo sapiens; RIP-Seq
Sample: SRS17679636
BioProject: PRJNA972979
BioSample: SAMN35102697
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: Overexpression of V5-tagged FLAG
rip antibody: V5-specific antibody
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579325 40464608 3034845600 864.1 FLAG_IP1.fastq.gz, SRR24579325, SRR24579325.lite SRA
SRX20362709 SRP437920 RIP-Seq SINGLE
GSM7359796: HEK293T cells, FLAG_IP_2; Homo sapiens; RIP-Seq
Sample: SRS17679637
BioProject: PRJNA972979
BioSample: SAMN35102696
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T
cell line: HEK293T
cell type: Human Embryonic Kidney
genotype: WT
treatment: Overexpression of V5-tagged FLAG
rip antibody: V5-specific antibody
geo_loc_name: missing
collection_date: missing
Original files (1)
HEK293T
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR24579324 37396927 2804769525 802.7 FLAG_IP2.fastq.gz, SRR24579324, SRR24579324.lite SRA

Linked Publications (1)

Data Files (40)

Accession File Name Stored Type Output Type Mapping Assembly Size Download
FLAG_IN1.fastq.gz RIP-Seq 928.9 MB link
FLAG_IN1.fastq.gz RIP-Seq 928.9 MB link
FLAG_IN2.fastq.gz RIP-Seq 1.0 GB link
FLAG_IN2.fastq.gz RIP-Seq 1.0 GB link
FLAG_IP1.fastq.gz RIP-Seq 864.1 MB link
FLAG_IP1.fastq.gz RIP-Seq 864.1 MB link
FLAG_IP2.fastq.gz RIP-Seq 802.7 MB link
FLAG_IP2.fastq.gz RIP-Seq 802.7 MB link
RTCA_IN1.fastq.gz RIP-Seq 824.8 MB link
RTCA_IN1.fastq.gz RIP-Seq 824.8 MB link
RTCA_IN2.fastq.gz RIP-Seq 808.7 MB link
RTCA_IN2.fastq.gz RIP-Seq 808.7 MB link
RTCA_IP1.fastq.gz RIP-Seq 816.4 MB link
RTCA_IP1.fastq.gz RIP-Seq 816.4 MB link
RTCA_IP2.fastq.gz RIP-Seq 858.7 MB link
RTCA_IP2.fastq.gz RIP-Seq 858.7 MB link
SCAF8_IN1.fastq.gz RIP-Seq 746.9 MB link
SCAF8_IN1.fastq.gz RIP-Seq 746.9 MB link
SCAF8_IN2.fastq.gz RIP-Seq 882.8 MB link
SCAF8_IN2.fastq.gz RIP-Seq 882.8 MB link
SCAF8_IP1.fastq.gz RIP-Seq 782.9 MB link
SCAF8_IP1.fastq.gz RIP-Seq 782.9 MB link
SCAF8_IP2.fastq.gz RIP-Seq 852.6 MB link
SCAF8_IP2.fastq.gz RIP-Seq 852.6 MB link
STAU2_IN1.fastq.gz RIP-Seq 805.7 MB link
STAU2_IN1.fastq.gz RIP-Seq 805.7 MB link
STAU2_IN2.fastq.gz RIP-Seq 902.6 MB link
STAU2_IN2.fastq.gz RIP-Seq 902.6 MB link
STAU2_IP1.fastq.gz RIP-Seq 925.0 MB link
STAU2_IP1.fastq.gz RIP-Seq 925.0 MB link
STAU2_IP2.fastq.gz RIP-Seq 876.9 MB link
STAU2_IP2.fastq.gz RIP-Seq 876.9 MB link
TRNAU1AP_IN1.fastq.gz RIP-Seq 652.9 MB link
TRNAU1AP_IN1.fastq.gz RIP-Seq 652.9 MB link
TRNAU1AP_IN2.fastq.gz RIP-Seq 493.3 MB link
TRNAU1AP_IN2.fastq.gz RIP-Seq 493.3 MB link
TRNAU1AP_IP1.fastq.gz RIP-Seq 514.1 MB link
TRNAU1AP_IP1.fastq.gz RIP-Seq 514.1 MB link
TRNAU1AP_IP2.fastq.gz RIP-Seq 819.3 MB link
TRNAU1AP_IP2.fastq.gz RIP-Seq 819.3 MB link