Sequencing and structure probing of long RNAs using MarathonRT

Reverse transcriptase (RT) enzymes are indispensable tools for interrogating diverse aspects of RNA metabolism and transcriptome composition. Due to the growing interest in sequence and structural complexity of long RNA molecules, processive RT enzymes are now required for preserving linkage and information content in mixed populations of transcripts, and the low-processivity RT enzymes that are commercially available cannot meet this need. MarathonRT is encoded within a eubacterial group II intron and it has been shown to efficiently copy highly structured long RNA molecules in a single pass.

In this work, Yale University researchers systematically characterize MarathonRT as a tool enzyme and optimize its performance in a variety of applications that include single cycle reverse-transcription of long RNAs, in-cell SHAPE-MaP using ultra-long amplicons and the detection of natural RNA base modifications. By diversifying MarathonRT reaction protocols, the researchers provide an upgraded suite of tools for cutting-edge RNA research and clinical application.