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| Status |
Public on Oct 24, 2022 |
| Title |
Transcriptome-wide profiling and quantification of N6-methyladenosine by enzyme-assisted adenosine deamination [probes_assay] |
| Organisms |
Escherichia coli; synthetic construct |
| Experiment type |
Other
|
| Summary |
We report evolved TadA-assisted N6-methyladenosine sequencing (eTAM-seq), an enzyme-assisted sequencing technology for quantitative, base-resolution profiling of m6A. eTAM-seq functions by global adenosine deamination, enabling detection of m6A as persistent A. We demonstrate adenosine-to-inosine (I) conversion rates up to 99% using a hyperactive TadA variant. With eTAM-seq, we profile and quantify m6A in the whole transcriptomes of HeLa cells and mouse embryonic stem cells (mESCs), with simultaneous deconvolution of the transcriptome and epitranscriptome. Further, we showcase deep sequencing-free, site-specific m6A quantification with as few as 10 cells, an input demand that is at least 4 orders of magnitude lower than existing methods. Collectively, eTAM-seq enables sensitive detection and faithful quantification of m6A with limited RNA input, representing a novel solution to deciphering the epitranscriptome.
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| Overall design |
Two synthetic RNA probes with and without TadA8.20 treatment. One E. coli tRNA with or without WTTadA or TadA8.20 treatment.
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| Contributor(s) |
Xiao Y, Liu S, Ge R, Wu Y, He C, Chen M, Tang W |
| Citation(s) |
36593412 |
| |
| Submission date |
Apr 19, 2022 |
| Last update date |
Dec 01, 2023 |
| Contact name |
Shun Liu |
| Organization name |
University of Chicago
|
| Department |
Chemistry
|
| Lab |
Chuan He Lab
|
| Street address |
929 E. 57th. Street
|
| City |
Chicago |
| State/province |
IL |
| ZIP/Postal code |
60637 |
| Country |
USA |
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| Platforms (2) |
| GPL16085 |
Illumina MiSeq (Escherichia coli) |
| GPL17769 |
Illumina MiSeq (synthetic construct) |
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| Samples (14)
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| This SubSeries is part of SuperSeries: |
| GSE201064 |
Transcriptome-wide profiling and quantification of N6-methyladenosine by enzyme-assisted adenosine deamination |
|
| Relations |
| BioProject |
PRJNA828168 |
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