An Infectious cDNA Clone of SARS-CoV-2

Area of Science:

• Virology
• Molecular Biology
• Infectious Diseases

Background:

• The COVID-19 pandemic, caused by SARS-CoV-2, highlights the need for robust experimental systems.
• Studying SARS-CoV-2 is crucial for developing effective countermeasures.

Purpose of the Study:

• To establish a reverse genetic system for SARS-CoV-2.
• To create an infectious clone of SARS-CoV-2 (icSARS-CoV-2).
• To develop a reporter virus for evaluating antiviral activities.

Main Methods:

• Assembly of seven cDNA fragments into a full-genome cDNA.
• Transcription of RNA from cDNA and electroporation into cells.
• Introduction of a reporter gene (mNeonGreen) into the viral genome.

Main Results:

• Highly infectious SARS-CoV-2 (icSARS-CoV-2) was produced with similar characteristics to clinical isolates.
• The engineered virus retained molecular markers and did not acquire mutations.
• A reporter virus (icSARS-CoV-2-mNG) was generated and used to evaluate interferon's antiviral activity.

Conclusions:

• The developed reverse genetic system provides a valuable tool for SARS-CoV-2 research.
• The reporter virus facilitates the study of viral replication and the evaluation of antiviral drugs.
• These reagents are key for advancing the development of countermeasures against SARS-CoV-2.