Single-molecule RNA sizing enables quantitative analysis of alternative transcription termination.

ABSTRACT: Transcription, a critical process in molecular biology, has found many applications in RNA synthesis, including mRNA vaccines and RNA therapeutics. However, current RNA characterization technologies suffer from amplification and enzymatic biases that lead to loss of native information. Here, we introduce a strategy to quantitatively study both transcription and RNA polymerase behaviour by sizing RNA with RNA nanotechnology and nanopores. To begin, we utilize T7 RNA polymerase to transcribe linear DNA lacking termination sequences. Surprisingly, we discover alternative transcription termination in the origin of replication sequence. Next, we employ circular DNA without transcription terminators to perform rolling circle transcription. This allows us to gain valuable insights into the proces

SUBMITTER: Patino-Guillen G 

PROVIDER: S-EPMC10894232 | biostudies-literature | 2024 Feb

REPOSITORIES: biostudies-literature