Systematic identification of RNA-binding proteins and tethered domains that activate exon splicing inclusion [eCLIP-seq]
Organism:
Homo sapiens
Platform:
GPL24676
Samples:
20
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
SRA Experiments (20) and Runs (20)
Total: 16188 MB
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579347 |
20833620 |
2104195620 |
652.87 |
TRNAU1AP_IN1.fastq.gz, SRR24579347, SRR24579347.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579346 |
15672441 |
1582916541 |
493.3 |
TRNAU1AP_IN2.fastq.gz, SRR24579346, SRR24579346.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579345 |
16806219 |
1697428119 |
514.11 |
TRNAU1AP_IP1.fastq.gz, SRR24579345, SRR24579345.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579344 |
26663497 |
2693013197 |
819.35 |
TRNAU1AP_IP2.fastq.gz, SRR24579344, SRR24579344.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579343 |
38684019 |
2901301425 |
824.83 |
RTCA_IN1.fastq.gz, SRR24579343, SRR24579343.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579342 |
37878702 |
2840902650 |
808.68 |
RTCA_IN2.fastq.gz, SRR24579342, SRR24579342.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579341 |
38085130 |
2856384750 |
816.45 |
RTCA_IP1.fastq.gz, SRR24579341, SRR24579341.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579340 |
40049683 |
3003726225 |
858.7 |
RTCA_IP2.fastq.gz, SRR24579340, SRR24579340.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579339 |
35063895 |
2629792125 |
746.9 |
SCAF8_IN1.fastq.gz, SRR24579339
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579338 |
41407652 |
3105573900 |
882.84 |
SCAF8_IN2.fastq.gz, SRR24579338, SRR24579338.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579337 |
36476437 |
2735732775 |
782.86 |
SCAF8_IP1.fastq.gz, SRR24579337, SRR24579337.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579336 |
39933385 |
2995003875 |
852.63 |
SCAF8_IP2.fastq.gz, SRR24579336, SRR24579336.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579335 |
37690833 |
2826812475 |
805.71 |
STAU2_IN1.fastq.gz, SRR24579335, SRR24579335.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579334 |
42305202 |
3172890150 |
902.56 |
STAU2_IN2.fastq.gz, SRR24579334, SRR24579334.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579333 |
43171073 |
3237830475 |
925.04 |
STAU2_IP1.fastq.gz, SRR24579333, SRR24579333.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579332 |
40871112 |
3065333400 |
876.95 |
STAU2_IP2.fastq.gz, SRR24579332, SRR24579332.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579327 |
43147058 |
3236029350 |
928.86 |
FLAG_IN1.fastq.gz, SRR24579327, SRR24579327.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579326 |
47732648 |
3579948600 |
1028.85 |
FLAG_IN2.fastq.gz, SRR24579326, SRR24579326.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579325 |
40464608 |
3034845600 |
864.1 |
FLAG_IP1.fastq.gz, SRR24579325, SRR24579325.lite
|
SRA
|
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)
Runs (1)
| Run |
Spots |
Bases |
Size (MB) |
Files |
Link |
| SRR24579324 |
37396927 |
2804769525 |
802.7 |
FLAG_IP2.fastq.gz, SRR24579324, SRR24579324.lite
|
SRA
|