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Status |
Public on Nov 13, 2020 |
Title |
MeRIP-Seq of pancreatic islets in Mettl3flox/flox and islet β cell-specific Mettl3 knockout (β-Mettl3-KO) mice |
Organism |
Mus musculus |
Experiment type |
Expression profiling by high throughput sequencing
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Summary |
Purpose: Through mRNA m6A-profiling, we aim to characterize the mRNA m6A changes in pancreatic islets obtained from Mettl3flox/flox and β-Mettl3-KO mice. Methods: Total RNA was extracted using Tripure Isolation Reagent (Roche, Mannheim, Germany) from pancreatic islets of Mettl3flox/flox and β-Mettl3-KO mice at 8 weeks old. A total of 2-3 μg RNAs were pooled from nine Mettl3flox/flox mice and twelve β-Mettl3-KO mice, respectively. Three independent biological replicates of each group were used for MeRIP-seq. Fragmented RNA (~100 nt) was incubated for 2 hr at 4oC with anti-m6A polyclonal antibody (Merk Millipore) in the immunoprecipitation experiment. Then, immunoprecipitated RNAs or Input was used for library construction with Ovation SoLo RNA-Seq System Core Kit (NuGEN). The library preparations were sequenced on an Illumina Novaseq platform with a paired-end read length of 150 bp according to the standard protocols. After mapping reads to the reference genome, exomePeak R package (version 2.16.0) was used for the m6A peak identification in each anti-m6A immunoprecipitation group with the corresponding input samples serving as a control, and q-value threshold of enrichment of 0.05 was used for all data sets. The m6A-enriched motifs of each group were identified by HOMER (version 4.9.1). Conclusion: The islet mRNA m6A profiles in Mettl3flox/flox and β-Mettl3-KO mice were characterized.
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Overall design |
To map the mRNA m6A modification caused by METTL3 in pancreatic islets, MeRIP-seq was performed in pancreatic islets obtained from Mettl3flox/flox and β-Mettl3-KO mice.
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Contributor(s) |
Chen Z, Li X |
Citation(s) |
33417895 |
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Submission date |
Aug 03, 2020 |
Last update date |
Feb 13, 2021 |
Contact name |
Zheng Chen |
E-mail(s) |
chenzheng@hit.edu.cn
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Phone |
+8645186402029
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Organization name |
Harbin Institute of Technology
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Street address |
92 Xidazhijie Harbin Institute of Technology
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City |
Harbin |
State/province |
Heilongjiang |
ZIP/Postal code |
150001 |
Country |
China |
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Platforms (1) |
GPL24247 |
Illumina NovaSeq 6000 (Mus musculus) |
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Samples (12)
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Relations |
BioProject |
PRJNA650499 |
SRA |
SRP275644 |