Transcription, catalyzed by RNA polymerases, is one of the most fundamental processes in living cells. Characterizing the enzymatic process or RNA generation for use in CRISPR requires quantifying RNA transcripts. This typically requires purification of the nucleic acids leading to experimental inaccuracies and loss of product. Synthetic fluorescent dyes are available but are relatively expensive. Here, we describe the use of a fluorescent biosensor based upon the single-stranded binding (SSB) protein, which has previously been established as an ssDNA biosensor. The protein is easily expressed in E. coli and can be used without prior purification of the RNA, thereby providing a low cost, easy to use alternative for measuring mRNA.
With the novel sensor transcription components will be identified faster and improve understanding of the complex nature of transcription. The CLARIOstar assisted in assay development and reliably quantifies mRNA with MDCC-SSB.