Identification of SNHG15-associated microRNAs in SNHG15 overexpression BT549 cells using miRNA-Seq

LncRNAs have been shown to act as molecular sponges for miRNAs, and hence functionally affect the activity of other RNA transcripts targeted by the sponged miRNAs. SNHG15 is an oncogenic lncRNA in a variety of cancers including breast cancer, lung cancer, and hepatocellular carcinoma. Overall design: In the present study, we used MS2 pulldown and miRNA-Seq to characterize microRNAs that associated with SNHG15 in SNHG15 overexpression BT549 cells. Cell lysates were prepared from cultured BT549-SNHG15-MS2 cells and then incubated with Amylose resin coated with MBP-MCP (MS2 coat protein) or Amylose resin at 4°C for 5 h with gentle rotation. After incubation, the supernatant was removed by a short centrifugation and the Amylose beads were washed three times in lysis buffer,the co-precipitated RNAs were eluted with 100 µl lysis buffer containing 20 mM maltose. miRNAs associated with SNHG15 were extracted and analyzed by miRNA-seq.