A CRISPR-Cas and Tat Peptide with Fluorescent RNA Aptamer System for Signal Amplification in RNA Imaging

We reported on an efficient RNA imaging strategy based on a CRISPR-Cas and Tat peptide with a fluorescent RNA aptamer (TRAP-tag). Using modified CRISPR-Cas RNA hairpin binding proteins fused with a Tat peptide array that recruits modified RNA aptamers, this simple and sensitive strategy is capable of visualizing endogenous RNA in cells with high precision and efficiency. In addition, the modular design of the CRISPR-TRAP-tag facilitates the substitution of sgRNAs, RNA hairpin binding proteins, and aptamers in order to optimize imaging quality and live cell affinity. With CRISPR-TRAP-tag, exogenous GCN4, endogenous mRNA MUC4, and lncRNA SatIII were distinctly visualized in single live cells.

Automated summary

We presented an efficient RNA imaging strategy based on CRISPR-Cas and Tat peptide with TRAP-tag. By fusing modified CRISPR-Cas RNA hairpin binding proteins with a Tat peptide array that recruits modified RNA aptamers, this strategy allows highly precise and efficient visualization of endogenous RNA in cells. The modular design of CRISPR-TRAP-tag enables the substitution of sgRNAs, RNA hairpin binding proteins, and aptamers to optimize imaging quality and live cell affinity. We successfully visualized exogenous GCN4, endogenous mRNA MUC4, and lncRNA SatIII in single live cells using CRISPR-TRAP-tag.