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RNA-Seq data of ALKBH5 and FTO double knockout HEK293T human cells
Egor A Smolin1, Andrey I Buyan1, Dmitry N Lyabin1
1Russian Academy of Sciences, Institute of Protein Research, Pushchino 142290, Russia.
Data in Brief
|May 6, 2022
Summary
Complete failure in N6-methyladenosine (m6A) demethylation by ALKBH5 and FTO impacts gene expression. This study reveals widespread changes in gene splicing and expression following double knockout of these key RNA demethylases.
Area of Science:
- Molecular Biology
- Epigenetics
- RNA Biology
Background:
- N6-methyladenosine (m6A) is a crucial mRNA modification regulating gene expression.
- RNA demethylases ALKBH5 and FTO are responsible for removing the m6A mark.
- These demethylases share similar substrate recognition, allowing partial functional redundancy.
Purpose of the Study:
- To investigate the global impact of complete m6A demethylation failure.
- To analyze changes in gene expression and alternative splicing upon simultaneous knockout of ALKBH5 and FTO.
- To understand the functional consequences of abrogated m6A epitranscriptomic regulation.
Main Methods:
- High-throughput RNA sequencing (RNA-Seq) was performed on ALKBH5 and FTO double knockout HEK293T cells and wild type controls.
- RNA libraries were sequenced using Illumina NextSeq 500.
- Bioinformatic analysis, including read mapping and differential expression analysis in R, was employed.
Main Results:
- The absence of ALKBH5 and FTO led to significant alterations in the transcriptome.
- 5871 differentially expressed genes were identified when comparing double knockout to wild type cells.
- 166 alternatively spliced genes were detected, indicating a substantial impact on mRNA processing.
Conclusions:
- Complete failure of m6A demethylation profoundly affects gene expression and alternative splicing.
- ALKBH5 and FTO play critical roles in maintaining normal mRNA homeostasis.
- The findings highlight the importance of the m6A epitranscriptomic layer in gene regulation.
