AUTHOR=Gan Ying , Li Aolin , Liu Jun , Wang Xiaofei , Zhang Zhenan , Li Qinhan , Ye Xiongjun , Yao Lin , Zhang Qian 

TITLE=m6A-mRNA Methylation Regulates Gene Expression and Programmable m6A Modification of Cellular RNAs With CRISPR-Cas13b in Renal Cell Carcinoma

JOURNAL=Frontiers in Genetics

VOLUME=12

YEAR=2022

URL=https://www.frontiersin.org/journals/genetics/articles/10.3389/fgene.2021.795611

DOI=10.3389/fgene.2021.795611

ISSN=1664-8021

ABSTRACT=<p><bold>Background:</bold> N<sup>6</sup>-methyladenosine (m<sup>6</sup>A) is the most extensive messenger RNA modification. Despite recent advances in the biological roles of m<sup>6</sup>A, its role in the development and progression of renal cell carcinoma (RCC) remains unclear.</p><p><bold>Methods:</bold> In this study, we gained the transcriptome-wide m<sup>6</sup>A profile and gene expression pattern in RCC and paired adjacent peritumoral tissues by meRIP-seq and RNA-seq. m<sup>6</sup>A modifications of mRNAs were validated by meRIP-qPCR in tissues, and targeted methylation or demethylation was validated by using a CRISPR-Cas13b-based tool in RCC cell lines.</p><p><bold>Results:</bold> Our findings showed that there were 13,805 m<sup>6</sup>A peaks among 5,568 coding gene transcripts (mRNAs) in adjacent tissues and 24,730 m<sup>6</sup>A peaks among 6,866 mRNAs in tumor tissues. Furthermore, m<sup>6</sup>A modification sites were usually located in the coding sequences (CDS), and some near the start and stop codons. Gene Ontology analysis revealed that coding genes had differential N<sup>6</sup>-methyladenosine sites and were enriched in kidney development and cancer-related signaling pathways. We also found that different levels of m<sup>6</sup>A modifications could regulate gene expression.</p><p><bold>Conclusion:</bold> In summary, our results provided evidence for studying the potential function of RNA m<sup>6</sup>A modification and m<sup>6</sup>A-mediated gene expression regulation in human RCC.</p>