Related Experiment Video
Updated: Nov 28, 2025
11:40
Simplified Reverse Genetics Method to Recover Recombinant Rotaviruses Expressing Reporter Proteins
Published on: April 17, 2020
9.1K
Generation of recombinant rotaviruses encoding a split NanoLuc peptide tag
Pimfhun Pannacha1, Yuta Kanai1, Takahiro Kawagishi1
1Department of Virology, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan.
Biochemical and Biophysical Research Communications
|November 30, 2020
Summary
Researchers developed novel reporter rotaviruses (RVs) using a split NanoLuc luciferase system. These HiBiT reporter RVs effectively track viral replication and assess antiviral drug efficacy.
Area of Science:
- Virology
- Molecular Biology
- Biotechnology
Background:
- Recombinant viruses with reporter proteins are crucial for studying viral replication and transmission.
- Split NanoLuc luciferase (NLuc) systems offer sensitive detection through complementary protein fragments.
Purpose of the Study:
- To generate stable reporter rotaviruses (RVs) using a split NLuc system.
- To validate the utility of these reporter RVs for monitoring viral replication and evaluating antiviral agents.
Main Methods:
- A split NLuc system (LgBiT and HiBiT fragments) was employed to create reporter RVs.
- An 11 amino acid HiBiT peptide tag was fused to the NSP1 protein of simian and human RVs.
- NLuc activity in infected cell lysates was measured to correlate with RV replication.
Main Results:
- NLuc activity in virus-infected cell lysates directly paralleled viral replication levels.
- The system successfully monitored the antiviral activity of neutralizing antibodies and reagents.
- Reductions in NLuc expression indicated effective inhibition of recombinant HiBiT reporter viruses.
Conclusions:
- The developed HiBiT reporter RV systems are powerful tools for studying the rotavirus life cycle and pathogenesis.
- These reporter RVs provide a robust platform for the discovery and development of novel antiviral drugs.
