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| Status |
Public on Dec 12, 2017 |
| Title |
Massively parallel reporter assay of 3’UTR sequences identifies in vivo rules for mRNA degradation |
| Organism |
Danio rerio |
| Experiment type |
Other
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| Summary |
The stability of mRNAs is regulated by signals within their sequences, but a systematic and predictive understanding of the underlying sequence rules remains elusive. Here, we introduce UTR-Seq, a combination of massively parallel reporter assays and regression models, to survey the dynamics of tens-of-thousands of 3’UTR sequences during early zebrafish embryogenesis. UTR-Seq revealed two temporal degradation programs: a maternally encoded early-onset program and a late-onset program that accelerated degradation after zygotic genome activation. Three signals regulated early-onset rates: stabilizing poly-U and UUAG sequences, and destabilizing GC-rich signals. Three signals explained late-onset degradation: miR-430 seeds, AU-rich sequences and Pumilio recognition sites. Sequence based regression models translated 3’UTRs into their unique decay patterns, and predicted the in vivo impact of sequence signals on mRNA stability. Their application led to the successful design of artificial 3’UTRs that conferred specific mRNA dynamics. UTR-Seq provides a general strategy to uncover the rules of RNA cis-regulation.
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| Overall design |
temporal expression profiles of reporter mRNAs from zebrafish embryos (7 replicates total) or oocytes (2 replicates total). A total of five embryo replicates were collected using pre-adenylated (A+) reporters in three separate experiments. In the first experiment, a sample was collected every hour between 1h to 8h, and at 10h. In the second experiment, a single sample was collected every hour between 1h to 10h, and split into two after RNA extraction to produce two technical replicates (techrep). In the third experiment, two separate samples were collected every two hours between 2h to 10h to produce two same-day biological replicates (biorep). A total of two replicates were collected using non-adenylated (A-) reporters in two separate experiments. In the first experiment, a sample was collected every hour between 1h to 8h, and at 10h. In the second experiment (rep), samples were collected at 1h, 2h, 3h, 4h (3 samples), 6h, 8h and 10h.
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| Contributor(s) |
Rabani M, Schier AF |
| Citation(s) |
29225039 |
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| Submission date |
Nov 08, 2017 |
| Last update date |
May 15, 2019 |
| Contact name |
Michal Rabani |
| E-mail(s) |
michal.rabani@mail.huji.ac.il
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| Phone |
026585705
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| Organization name |
Hebrew University of Jerusalem
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| Street address |
Hebrew University, Edmond J. Safra Campus, Silberman building
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| City |
Jerusalem |
| ZIP/Postal code |
9190401 |
| Country |
Israel |
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| Platforms (2) |
| GPL18413 |
Illumina HiSeq 2500 (Danio rerio) |
| GPL21930 |
Illumina MiSeq (Danio rerio) |
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| Samples (71)
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| Relations |
| BioProject |
PRJNA417597 |
| SRA |
SRP124609 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE106677_seq0312.40A.1.txt.gz |
870.3 Kb |
(ftp)(http) |
TXT |
| GSE106677_seq0312.40A.2.txt.gz |
868.0 Kb |
(ftp)(http) |
TXT |
| GSE106677_seq0417.00A.txt.gz |
1.5 Mb |
(ftp)(http) |
TXT |
| GSE106677_seq0514.00A.txt.gz |
625.6 Kb |
(ftp)(http) |
TXT |
| GSE106677_seq0514.40A.txt.gz |
627.3 Kb |
(ftp)(http) |
TXT |
| GSE106677_seq0714.40A.1.txt.gz |
1.6 Mb |
(ftp)(http) |
TXT |
| GSE106677_seq0714.40A.2.txt.gz |
1.6 Mb |
(ftp)(http) |
TXT |
| GSE106677_seq1022.00A.txt.gz |
1.6 Mb |
(ftp)(http) |
TXT |
| GSE106677_seq1022.40A.txt.gz |
1.7 Mb |
(ftp)(http) |
TXT |
| GSE106677_sequences.txt.gz |
3.3 Mb |
(ftp)(http) |
TXT |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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