Department of Chemistry, The University of Chicago, Chicago, IL, USA.
Howard Hughes Medical Institute, The University of Chicago, Chicago, IL, USA.
Methods Mol Biol. 2021;2298:97-104. doi: 10.1007/978-1-0716-1374-0_6.
mG-seq detects internal 7-methylguanosine (mG) sites within mRNAs and noncoding RNAs by misincorporation signatures. A chemical-assisted sequencing approach selectively converts internal mG sites into abasic sites, triggering misincorporation at these sites in the presence of a specific reverse transcriptase. The further enrichment of mG-induced abasic sites by biotin pull-down reveals hundreds of internal mG sites in human mRNA. The misincorporation ratio before pull-down enrichment can be used for estimating the methylation fraction of some highly methylated mG sites.
mG-seq 通过错配特征检测 mRNA 和非编码 RNA 内的 7-甲基鸟苷(mG)位点。一种化学辅助测序方法选择性地将内部 mG 位点转化为无碱基位点，在特定逆转录酶存在的情况下，在这些位点引发错配。通过生物素下拉进一步富集 mG 诱导的无碱基位点，揭示了数百个人 mRNA 中的内部 mG 位点。下拉富集前的错配率可用于估计一些高度甲基化 mG 位点的甲基化分数。
