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A Seamless Cloning Approach for Porcine Reproductive and Respiratory Syndrome Virus Expression Vector Construction
Published on: May 17, 2024
Suppression of porcine arterivirus replication by baculovirus-delivered shRNA targeting nucleoprotein
Liqun Lu1, Yuenfern Ho, Jimmy Kwang
1Animal Health Biotechnology Unit, Temasek Life Sciences Laboratory, 1 Research Link, National University of Singapore, Singapore 117604, Singapore.
Abstract:
The ability to synthesize shRNAs from DNA templates driven by RNA polymerase III promoters has made it possible to apply virus-derived vectors as delivery vehicles for double-strand RNA-mediated interference. Baculovirus emerges as a promising vector for in vivo gene therapy most recently. To investigate its potential as a delivery vector for anti-virus shRNA targeting arterivirus porcine reproductive and respiratory syndrome virus (PRRSV), we constructed recombinant baculovirus vectors bearing a shRNA-synthesizing cassette driven by U6 promoter with enhanced transduction efficiency by displaying vesicular stomatitis virus glycoprotein on viral envelope. Transduction of Marc145 cells with a recombinant baculovirus delivering egfp gene-specific shRNA dramatically suppressed the expression of EGFP in this cell line; and transduction of Marc145 cells with a baculovirus delivering shRNA specific for the C-terminal nucleoprotein coding region of PRRSV genome resulted in inhibition of viral replication. Our data highlight the recombinant baculovirus as an alternative vehicle for anti-virus shRNA delivery.
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