Engineering efficient termination of bacteriophage T7 RNA polymerase transcription.

ABSTRACT: The bacteriophage T7 expression system is one of the most prominent transcription systems used in biotechnology and molecular-level research. However, T7 RNA polymerase is prone to read-through transcription due to its high processivity. As a consequence, enforcing efficient transcriptional termination is difficult. The termination hairpin found natively in the T7 genome is adapted to be inefficient, exhibiting 62% termination efficiency in vivo and even lower efficiency in vitro. In this study, we engineered a series of sequences that outperform the efficiency of the native terminator hairpin. By embedding a previously discovered 8-nucleotide T7 polymerase pause sequence within a synthetic hairpin sequence, we observed in vivo termination efficiency of 91%; by joining 2 short sequences in

SUBMITTER: Calvopina-Chavez DG 

PROVIDER: S-EPMC9157156 | biostudies-literature | 2022 May

REPOSITORIES: biostudies-literature