fox files

Raising national security concerns.

A Thursday npj Vaccines study confirms U.S. federal agencies, including the U.S. Department of Agriculture (USDA) and the National Institutes of Health (NIH), funded the laboratory creation of genetically engineered two influenza viruses (H9N2/H5N2) built from plasmids, artificial gene fusions, synthetic insertions, and modified genome segments.

The work was carried out by researchers at the University of Georgia, the Icahn School of Medicine at Mount Sinai, and the U.S. National Poultry Research Center, Agricultural Research Service, USDA.

The authors listed on the study are: “Flavio Cargnin Faccin, L. Claire Gay, Dikshya Regmi, Robert Hoelzl, Teresa D. Mejías, Darrell Kapczynski, Florian Krammer & Daniel R. Perez.”

The study’s funding confirms direct federal involvement.

“Funding for this work includes grants… National Institute of Food and Agriculture (NIFA), U.S. Department of Agriculture (USDA) Grant award numbers 2021-67015-33406 and 2024-67015-42736, National Institute of Allergy and Infectious Diseases, National Institutes of Health (NIH) Contract number 75N93021C00014.”

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Construction of the Engineered Viruses

The researchers did not isolate a purported virus from nature.

They built new viruses entirely through reverse genetics, using plasmid DNA transfected into human and dog cell lines.

The paper states: “Recombinant viruses were rescued by reverse genetics using the 8-plasmid system and helper plasmids in a coculture of HEK293T and MDCK cells.”

The process is described explicitly.

According to the authors, “1 µg of each plasmid was mixed… and used to overlay the cell coculture.”

Viral stocks were then expanded artificially rather than occurring naturally, as the study reports: “Viral stocks were generated in 10-day-old specific pathogen-free (SPF) eggs.”

These details confirm that the viruses were constructed in the laboratory, rescued from DNA, and amplified in eggs.

‘Chimeric’ Pathogens

The viruses created under USDA and NIH funding are laboratory-assembled chimeras—genomes stitched together from engineered parts that do not exist in nature.

The study shows that two influenza proteins were fused into a single artificial gene, something no wild virus carries.

As the authors write: “Segment 2 was modified to encode a chimeric PB1-M2 open reading frame (ORF) separated by a glycine-glycine-glycine-glycine-serine (G4S) spacer.”

This forces the virus to make an unnatural hybrid protein.

To ensure the virus depends on this man-made fusion protein, its normal version of M2 was deliberately shut off.

The paper states: “Segment 7 was modified by introducing multiple early stop codons in the M2 ORF via site-directed mutagenesis to prevent its expression.”

This eliminates the native M2 and locks the virus into the engineered design.

The researchers also inserted a synthetic 58–amino acid sequence into the HA segment, including an artificial peptide not found in any influenza strain.

The methods describe this as: “Segment 4… was modified to insert a 58-amino-acid-long sequence… which included the unique 8-amino-acid peptide… ‘DRPAVIAN.’”

Another modification swaps out the cleavage site of an H5 virus and replaces it with one taken from a human 1934 H1N1 strain, altering how the virus activates inside host cells.

The authors state: “The H5 HA HPAI cleavage site was replaced with that of the A/Puerto Rico/8/1934 (H1N1) (PR8) strain.”

Finally, the virus was engineered to manufacture a chicken immune-signaling molecule from inside infected cells.

The study confirms: “The mature protein-coding sequence of chicken IL-18… was subcloned in frame with the NA ORF.”

These combined changes—fusion genes, disabled native proteins, synthetic inserts, human-strain cleavage sites, and cytokine-expression modules—create a virus with properties that no purported natural influenza lineage carries.

Aerosol Exposure in Animals

The researchers then exposed day-old chickens to the engineered viruses through aerosolized live-virus delivery.

The methods describe this process clearly, all done within BSL-3 lab conditions:

“One-day old SPF White Leghorn chickens… were vaccinated via aerosol using an aerosol chamber… A 5 mL volume of MLV-H9N2-IL was loaded into the Aeroneb lab nebulizer, resulting in an average exposure of 1×10⁶ EID50/chicken… The exposure lasted for 15 min.”

This confirms that the USDA- and NIH-funded engineered viruses were not only constructed but also introduced into animals through airborne delivery.

Bottom Line

The study documents how U.S. federal agencies oversaw the full laboratory assembly of engineered influenza viruses—built from plasmids, redesigned through reverse genetics, and altered with fusion genes, stop-codon knockouts, synthetic peptide insertions, cytokine-expression modules, and foreign cleavage sites.

These are not environmental isolates; they are fully man-made constructs created inside U.S. government and university laboratories under USDA and NIH funding.

The viruses were then delivered to live animals by aerosol, demonstrating not only construction capability but functional deployment.

Work of this nature carries obvious national-security implications: it establishes the technical capacity to design, modify, and disseminate engineered influenza strains whose properties cannot be predicted from any purported natural lineage.