Promoter RNA sequencing (PRSeq) for the massive and quantitative promoter analysis in vitro.

ABSTRACT: Analysis of promoter strength and specificity is important for understanding and engineering gene regulation. Here, we report an in vitro promoter analysis method that can achieve both massiveness and quantitativeness. In this approach, a pool of single-stranded DNA with a partially randomized promoter sequence to be analyzed is chemically synthesized. Through enzymatic reactions, the randomized sequence will be copied to the downstream region, resulting in a template DNA pool that carries its own promoter information on its transcribed region. After in vitro transcription of the DNA pool with an RNA polymerase of interest, the sequences of the resulting transcripts will be analyzed. Since the promoter strength linearly correlates to the copy number of transcript, the strength of each prom

SUBMITTER: Ohuchi S 

PROVIDER: S-EPMC6395800 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature