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An infectious cDNA clone of a growth attenuated Korean isolate of MERS coronavirus KNIH002 in clade B

Minwoo Kim1, Hee Cho1, Seung-Hoon Lee1

  • 1Department of Biotechnology, Yonsei University, Seoul, Korea.

Emerging Microbes & Infections
|December 9, 2020
PubMed

Insights

A new reverse genetics system for Middle East respiratory syndrome-coronavirus (MERS-CoV) clade B (KNIH002) was developed. This system revealed MERS-CoV genetic instability and growth attenuation, crucial for future MERS-CoV research.

Area of Science:

  • Virology
  • Molecular Biology
  • Epidemiology

Background:

  • The 2015 Korean MERS-CoV outbreak involved a clade B strain (KNIH002) with distinct characteristics from the clade A prototype (EMC strain).
  • Understanding MERS-CoV evolution and pathophysiology is crucial for public health.

Purpose of the Study:

  • To establish a BAC-based reverse genetics system for the MERS-CoV KNIH002 strain.
  • To investigate the genetic stability and growth characteristics of a recombinant MERS-CoV derived from the KNIH002 strain.
  • To facilitate comparative studies between MERS-CoV clades for better understanding of its pathophysiology.

Main Methods:

  • Development of a Bacterial Artificial Chromosome (BAC)-based reverse genetics system.
  • Generation of a recombinant MERS-CoV expressing red fluorescent protein from the KNIH002 isolate.
  • Comparative analysis of viral growth and genetic stability with the MERS-CoV EMC strain.

Main Results:

  • The rescued recombinant MERS-CoV and the KNIH002 strain exhibited growth attenuation compared to the EMC strain.
  • Rapid generation of ORF5 variants upon serial passage of the rescued virus, indicating genetic instability.
  • The infectious clone provides a tool for comparative MERS-CoV studies.

Conclusions:

  • The developed infectious clone for MERS-CoV clade B (KNIH002) is a valuable tool for studying MERS-CoV evolution and pathophysiology.
  • The genetic instability observed highlights the need for caution when using passaged viruses in research.
  • Comparative studies using this system can elucidate the functional link between MERS-CoV molecular evolution and disease.
Keywords:
MERS-CoVORF5 deletion variantsclade B Korean isolategrowth attenuationinfectious cDNA clone

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