tetano
Editor, Senior Moderator
Proc Natl Acad Sci U S A
. 2026 Jun 30;123(26):e2532920123.
doi: 10.1073/pnas.2532920123. Epub 2026 Jun 24.
Extra gene coding capacity of SARS-CoV-2 provides a virus engineering platform for in vitro and in vivo applications
Taksoo Kim[SUP] 1 [/SUP], Asim Biswas[SUP] 1 [/SUP], Sangam Kandel[SUP] 1 [/SUP], Makoto Kuroda[SUP] 1 [/SUP], Peter J Halfmann[SUP] 1 [/SUP], Yoshihiro Kawaoka[SUP] 1 2 3 4 [/SUP]
Affiliations
The genomic flexibility of orthocoronaviruses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is showcased by the presence of accessory genes, which vary in number among virus species and strains. Given this flexibility, the viral coding capacity can be artificially expanded to express a heterologous gene from the viral genome, thereby enabling the development of a viral vector platform. Here, we systematically explored the extra gene coding capacity of SARS-CoV-2 by inserting an extra reporter gene at every intergenic region in its genome. We revealed the entire scheme of its extra gene expression and identified a genomic location that stably expresses reporter genes while maintaining the wild-type viral phenotype. Using this construct, we developed a set of fluorescent and luminescent reporter SARS-CoV-2 viruses available for in vivo flow cytometry and in vitro antiviral screening. Flow cytometric analysis with these reporter viruses revealed cell type-specific dynamics of SARS-CoV-2 infection in the lung tissue of K18-hACE2 mice. Our findings offer a platform for SARS-CoV-2 genome engineering, providing a set of reporter viruses for research applications.
Keywords: SARS-CoV-2; coronavirus; in vivo flow cytometry; reporter virus; viral vector.
. 2026 Jun 30;123(26):e2532920123.
doi: 10.1073/pnas.2532920123. Epub 2026 Jun 24.
Extra gene coding capacity of SARS-CoV-2 provides a virus engineering platform for in vitro and in vivo applications
Taksoo Kim[SUP] 1 [/SUP], Asim Biswas[SUP] 1 [/SUP], Sangam Kandel[SUP] 1 [/SUP], Makoto Kuroda[SUP] 1 [/SUP], Peter J Halfmann[SUP] 1 [/SUP], Yoshihiro Kawaoka[SUP] 1 2 3 4 [/SUP]
Affiliations
- PMID: 42341043
- DOI: 10.1073/pnas.2532920123
The genomic flexibility of orthocoronaviruses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is showcased by the presence of accessory genes, which vary in number among virus species and strains. Given this flexibility, the viral coding capacity can be artificially expanded to express a heterologous gene from the viral genome, thereby enabling the development of a viral vector platform. Here, we systematically explored the extra gene coding capacity of SARS-CoV-2 by inserting an extra reporter gene at every intergenic region in its genome. We revealed the entire scheme of its extra gene expression and identified a genomic location that stably expresses reporter genes while maintaining the wild-type viral phenotype. Using this construct, we developed a set of fluorescent and luminescent reporter SARS-CoV-2 viruses available for in vivo flow cytometry and in vitro antiviral screening. Flow cytometric analysis with these reporter viruses revealed cell type-specific dynamics of SARS-CoV-2 infection in the lung tissue of K18-hACE2 mice. Our findings offer a platform for SARS-CoV-2 genome engineering, providing a set of reporter viruses for research applications.
Keywords: SARS-CoV-2; coronavirus; in vivo flow cytometry; reporter virus; viral vector.