Targeted point mutations of the m6A modification in miR675 using RNA-guided base editing induce cell apoptosis
| Autor: | Yidan Xia, Dongxu Wang, Chengshun Li, Ziping Jiang, Chao Lin, Yang Hao, Jindong Hao, Xianfeng Yu, Fei Gao |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Biophysics Apoptosis Methylation Biochemistry m6A modification 03 medical and health sciences 0302 clinical medicine microRNA Gene expression Humans Point Mutation Epigenetics RNA Processing Post-Transcriptional Molecular Biology Research Articles Cancer Gene Editing Gene knockdown miR675 Gene Expression & Regulation cell apoptosis H19 Chemistry Adenine Point mutation HEK 293 cells RNA Cell Biology Cell biology MicroRNAs HEK293 Cells 030104 developmental biology Gene Expression Regulation A549 Cells 030220 oncology & carcinogenesis gene expression RNA Long Noncoding CRISPR-Cas Systems RNA Guide Kinetoplastida |
| Zdroj: | Bioscience Reports |
| ISSN: | 1573-4935 0144-8463 |
| DOI: | 10.1042/bsr20192933 |
| Popis: | Methylation of the adenine base at the nitrogen 6 position (m6A) is the most common post-transcriptional epigenetic modification of RNA, and it plays a very important role in regulating gene expression. To investigate the role of m6A methylation in the expression of non-coding RNA and miRNA, we used a system of adenine base editors (ABEs). Here, we mutated regions up- and downstream of miRNA 675 m6A modification sites in the H19 locus using HEK293T, L02, MHCC97L, MHCC97H, A549, and SGC-7901 cells. Our results showed that a T–A base transversion had occurred in all cell lines. Moreover, mutation of the regions upstream of the miRNA 675 m6A modification site led to reduced expression of H19 and the induction of cell apoptosis in HEK293T cells. To further confirm our results, L02 and MHCC97L cells were detected using ABEs system. The results indicated increased cell apoptosis and reduced expression of miR675 as well as H19. To confirm the relationship between H19 and miR675 expression, overexpression and knockdown studies were performed. The results showed that reduced HI9 expression induced cell apoptosis through miR675. Taken together, these results indicate that m6A modification can regulate the expression of H19 and miR675 which induce cell apoptosis. |
| Databáze: | OpenAIRE |
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