Intelligent Guide RNA: Harboring Dual Toe-Hold Switches for Luciferase Reporter Modulation in the Presence of Trigger RNA

Abstract

Abstract The CRISPR system has been extensively utilized in various aspects of molecular biology for over a decade. However, its continuous activity can result in undesirable effects. Nevertheless, implementing programmable CRISPR/Cas9 function using nano-device mediation can effectively mitigate these side effects. By integrating RNA-sensing platforms, into the CRISPR system, it has been transformed into a powerful tool for processing internal cell information and modulating gene activity. Here, an Intelligent guide RNA (IngRNA), a cis-repressed gRNA synthetic circuit which enables efficient recognition of arbitrary stimulatory RNA was developed. In this system, the presence of cognate trigger RNA promotes precise binding to the first toe-hold site, subsequently exposes the second toe-hold site. The connection between the 5' end of platform and the second toe-hold site facilitates gRNA activity to target gene of interest. Activating this genetic logic circuit in response to specific cellular conditions holds great promise as a platform for synthetic biology.