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| Status |
Public on Apr 10, 2024 |
| Title |
Detection of internal m7G modification in the human transcriptome in a quantitative manner and at single-nucleotide resolution |
| Organism |
Homo sapiens |
| Experiment type |
Non-coding RNA profiling by high throughput sequencing
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| Summary |
RNA modifications play an important role in regulating RNA stability and gene expression, however the development of reliable detection techniques has been a major burden in the field. Here, we describe two different methodologies for global identification of N-7-methylguanosine (m7G) in human tRNAs. An antibody and northdot blot based quantitative approach that allows global detection of m7G levels. And borohydride reduction sequencing (Bo-Seq), a chemically based approach that maps m7G modification at single nucleotide resolution. The specificity of Bo-Seq lies on RNA size-selection, optimized RNA scission using NaBH4, aniline and m7GTPs, and elimination of complex chemical steps or bioinformatic analysis included in similar methods, and reduces the protocol to four days. Bo-Seq and our antibody based approach are specifically developed for easily and globally detect tRNA m7G-methylation in human cells. We also validated METTL1 as the methyltransferase that methylates tRNAs in human cells.
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| Overall design |
Small RNA from prostate cancer cells DU145 were extrated. RNA were either non chemically treated or treated wih NaBH4 and aniline (BH4). Two replicates were used per condition. A total of 4 samples were sequenced.
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| Contributor(s) |
Blanco S, D'Ambrosi S, García-Vílchez R, Dietmann S, Aransay AM |
| Citation(s) |
38566310 |
| |
| Submission date |
May 18, 2022 |
| Last update date |
Apr 10, 2024 |
| Contact name |
Ana Maria Aransay |
| E-mail(s) |
amaransay@cicbiogune.es
|
| Phone |
0034944061325
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| Organization name |
CIC bioGUNE
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| Department |
Genome Analysis Platform
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| Street address |
Parque tecnologico de Bizkaia, Building 801-A
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| City |
Derio |
| State/province |
BIZKAIA |
| ZIP/Postal code |
48160 |
| Country |
Spain |
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| Platforms (1) |
| GPL16791 |
Illumina HiSeq 2500 (Homo sapiens) |
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| Samples (4)
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| Relations |
| BioProject |
PRJNA839265 |
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