Evaluation of Virus-Free Manufacture of Recombinant Proteins Using CRISPR-Mediated Gene Disruption in Baculovirus-Infected Insect Cells.
Mark R BruderMarc G AucoinPublished in: Vaccines (2023)
The manufacture and downstream processing of virus-like particles (VLPs) using the baculovirus expression vector system (BEVS) is complicated by the presence of large concentrations of baculovirus particles, which are similar in size and density to VLPs, and consequently are difficult to separate. To reduce the burden of downstream processing, CRISPR-Cas9 technology was used to introduce insertion-deletion (indel) mutations within the Autographa californica multiple nucleopolyhedrovirus ( Ac MNPV) gp64 open reading frame, which encodes the major envelope protein of Ac MNPV. After comfirming the site-specific targeting of gp64 leading to reduced budded virus (BV) release, the gag gene of human immunodeficiency virus type 1 was expressed to produce Gag VLPs. This approach was effective for producing VLPs using the BEVS whilst simultaneously obstructing BV release.
Keyphrases
- human immunodeficiency virus
- crispr cas
- genome editing
- genome wide
- hepatitis c virus
- antiretroviral therapy
- lipopolysaccharide induced
- copy number
- induced apoptosis
- poor prognosis
- hiv infected
- binding protein
- minimally invasive
- dna methylation
- hiv aids
- disease virus
- transcription factor
- protein protein
- endoplasmic reticulum stress
- oxidative stress
- gene expression