Determine the methyltransferase activities of METTL16 via MeRIP-seq (m6A-seq)

METTL16 belongs to methyltransferase like (METTL) family and could install m6A marks on its substrates. Here, we uncover a tumor-promoting role of METTL16 in AML and LSC self-renewal. To explore the mechanism underlying the oncogenic function of METTL16 in AML, we performed m6A-seq and RNA-seq. Via integrated analysis of m6A-seq data and RNA-seq data, we identified two bona fide targets of METTL16, BCAT1 and BCAT2. METTL16 functions as an m6A methyltransferase to regulate expression of BCAT1 and BCAT2, which contribute to reprogramming of BCAA metabolism. Overall design: The NOMO-1 cells were infected with sgMETTL16-1, sgMETTL16-2,and sgNS lentivirus to induce knockout of METTL16. The positive-infected cells were selected with 2ug/ml puromycin for 1-2 weeks. The cells were collected and total RNA were isolated from the whole cells via Qiazol. The poly(A) RNAs were further enriched from the total RNA and then subjected to m6A IP.