text: miCLIP (Methylation induced crosslinking immunoprecipitation) was used to detect NSUN2 targets, which were found to be mostly non-coding RNAs such as tRNA. An induced mutation of C271A in NSUN2 inhibits release of enzyme from RNA target. This mutation was over-expressed in the cells of interest, and the mutated NSUN2 was also tagged with the Myc epitope. The covalently linked RNA-protein complexes are isolated via immunoprecipitation for a Myc-specific antibody. These complexes are confirmed and detected by radiolabeling with phosphorus-32. The RNA is then extracted from the complex, reverse-transcribed, amplified with PCR, and sequenced using next-generation platforms.
  Both miCLIP and Aza-IP, though limited by specific targeting of enzymes, can allow for the detection of low-abundance methylated RNA without deep sequencing.