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| Status |
Public on Mar 30, 2023 |
| Title |
BuTT-Seq: a new method for facile profiling of transcription |
| Organism |
Drosophila melanogaster |
| Experiment type |
Other
Expression profiling by high throughput sequencing
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| Summary |
A wide range of sequencing methods have been developed to assess nascent RNA transcription and resolve the single-nucleotide position of RNA polymerase genome-wide. These techniques are often burdened with high input material requirements and lengthy protocols. We leveraged the template-switching properties of thermostable group II intron reverse transcriptase (TGIRT) and developed BuTT-Seq (BUlk analysis of nascent Transcript Termini sequencing), which can produce libraries from purified nascent RNA in 6 hours and from as few as 10,000 cells – an improvement of at least 10-fold over existing techniques. BuTT-Seq shows that inhibition of the superelongation complex (SEC) causes promoter-proximal pausing to move upstream in a fashion correlated with subnucleosomal fragments. To address transcriptional regulation in a tissue, BuTT-Seq was used to measure the circadian regulation of transcription from fly heads. All the results indicate that BuTT-Seq is a simple and powerful technique to analyze transcription at a high level of resolution.
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| Overall design |
Comparing transcriptional pausing and elongation with or without treatment with Superelongation Complex inhibitor in S2 Cells as well as assaying Circadian transcription around the clock in Drosophila.
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| Contributor(s) |
Yu AD, Rosbash M |
| Citation(s) |
37164645 |
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| Submission date |
Mar 30, 2023 |
| Last update date |
Sep 08, 2023 |
| Contact name |
Albert Dayuan Yu |
| E-mail(s) |
adyu@brandeis.edu
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| Phone |
8579917778
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| Organization name |
Brandeis University
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| Department |
Molecular and Cell Biology
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| Lab |
Rosbash Lab
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| Street address |
415 South St.
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| City |
Waltham |
| State/province |
MA |
| ZIP/Postal code |
02453 |
| Country |
USA |
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| Platforms (1) |
| GPL19132 |
Illumina NextSeq 500 (Drosophila melanogaster) |
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| Samples (19)
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| GSM7133672 |
DMSO Treated S2 Cells, BuTT-Seq, Replicate 1 |
| GSM7133673 |
DMSO Treated S2 Cells, BuTT-Seq, Replicate 2 |
| GSM7133674 |
KL-1 Treated S2 Cells, BuTT-Seq, Replicate 1 |
| GSM7133675 |
KL-1 Treated S2 Cells, BuTT-Seq, Replicate 2 |
| GSM7133676 |
CantonS Heads, ZT2, Replicate 1 |
| GSM7133677 |
CantonS Heads, ZT2, Replicate 2 |
| GSM7133678 |
CantonS Heads, ZT6, Replicate 1 |
| GSM7133679 |
CantonS Heads, ZT6, Replicate 2 |
| GSM7133680 |
CantonS Heads, ZT10, Replicate 1 |
| GSM7133681 |
CantonS Heads, ZT10, Replicate 2 |
| GSM7133682 |
CantonS Heads, ZT14, Replicate 1 |
| GSM7133683 |
CantonS Heads, ZT14, Replicate 2 |
| GSM7133684 |
CantonS Heads, ZT18, Replicate 1 |
| GSM7133685 |
CantonS Heads, ZT18, Replicate 2 |
| GSM7133686 |
CantonS Heads, ZT22, Replicate 1 |
| GSM7133687 |
CantonS Heads, ZT22, Replicate 2 |
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| Relations |
| BioProject |
PRJNA950415 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE228595_RAW.tar |
400.7 Mb |
(http)(custom) |
TAR (of BW) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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