A split ribozyme that links detection of a native RNA to orthogonal protein outputs

Individual RNA remains a challenging signal to synthetically transduce into different types of cellular information. To address this challenge we have developed Ribozyme-ENabled Detection of RNA (RENDR), a plug-and-play strategy that uses cellular transcripts to template the assembly of split ribozymes, triggering splicing reactions that generate orthogonal protein outputs. To identify split ribozymes that require templating for splicing, we used in vitro transposon mutagenesis to generate a library of split variants of the Tetrahymena thermophila ribozyme. We then screened the library using a fluorescence-activated cell sorting (FACS) to enrich for variants that were capable of complementation. Here we have deposited NGS data of the library of variants before and after sorting, as well as variants that were manually screened using bulk fluorescence data.