# Establishment of a Dual-Reporter Minigenome System for Respiratory Syncytial Virus

## TL;DR

A new dual-reporter system for Respiratory Syncytial Virus helps screen antiviral drugs more effectively and safely.

## Contribution

The development of a dual-reporter minigenome system for RSV using NanoLuc and sfGFP for antiviral screening.

## Key findings

- The dual-reporter system reliably detects antiviral activity using orthogonal signals from NanoLuc and sfGFP.
- Validation with L polymerase inhibitors confirmed the system's effectiveness for small-molecule screening.
- The system shows a linear correlation between reporter signals, enhancing assay reliability.

## Abstract

Respiratory syncytial virus (RSV) poses a significant global health challenge, particularly affecting infants, the elderly, and immunocompromised individuals. Despite recent progress in the development of vaccines and monoclonal antibodies, effective antiviral therapies remain limited. To advance the discovery of antiviral drugs, we have developed a dual-reporter RSV minigenome system, providing a safe and robust platform for antiviral evaluation. This system incorporates NanoLuc luciferase and superfolder GFP (sfGFP) linked by a self-cleaving P2A peptide, allowing for the simultaneous detection of orthogonal signals. Validation with L polymerase inhibitors confirmed the system’s reliability for screening small-molecule inhibitors. The linear correlation observed between the reporter signals enhances the assay’s reliability for antiviral assessment. This dual-reporter minigenome system advances targeted therapeutic strategies against RSV.