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Efficient production of inhibitor-free foamy virus glycoprotein-containing retroviral vectors by proteoglycan-deficient packaging cells

Foamy viruses (FVs) or heterologous retroviruses pseudotyped with FV glycoprotein enable transduction of a great variety of target tissues of disparate species. Specific cellular entry receptors responsible for this exceptionally broad tropism await their identification. Though, ubiquitously expressed heparan sulfate proteoglycan (HS-PG) is known to serve as an attachment factor of FV envelope (Env)-containing virus particles, greatly enhancing target cell permissiveness. Production of high-titer, FV Env-containing retroviral vectors is strongly dependent on the use of cationic polymer-based transfection reagents like polyethyleneimine (PEI). We identified packaging cell-surface HS-PG expression to be responsible for this requirement. Efficient release of FV Env-containing virus particles necessitates neutralization of HS-PG binding sites by PEI. Remarkably, remnants of PEI in FV Env-containing vector supernatants, which are not easily removable, negatively impact target cell transduction, in particular those of myeloid and lymphoid origin. To overcome this limitation for production of FV Env-containing retrovirus supernatants, we generated 293T-based packaging cell lines devoid of HS-PG by genome engineering. This enabled, for the first, time production of inhibitor-free, high-titer FV Env-containing virus supernatants by non-cationic polymer-mediated transfection. Depending on the type of virus, produced titers were 2- to 10-fold higher compared with those obtained by PEI transfection.

Identiferoai:union.ndltd.org:DRESDEN/oai:qucosa:de:qucosa:91679
Date04 June 2024
CreatorsMunz, Clara Marie, Kreher, Henriette, Erdbeer, Alexander, Stanke, Nicole, Richter, Stefanie, Westphal, Dana, Yi, Buqing, Behrendt, Rayk, Lindel, Fabian, Lindemann, Dirk
PublisherElsevier
Source SetsHochschulschriftenserver (HSSS) der SLUB Dresden
LanguageEnglish
Detected LanguageEnglish
Typeinfo:eu-repo/semantics/publishedVersion, doc-type:article, info:eu-repo/semantics/article, doc-type:Text
Rightsinfo:eu-repo/semantics/openAccess
Relation2329-0501, 10.1016/j.omtm.2022.07.004, info:eu-repo/grantAgreement/Deutsche Forschungsgemeinschaft/SPP 1923: Innate Sensing and Restriction of Retroviruses/273898015/, info:eu-repo/grantAgreement/Deutsche Forschungsgemeinschaft/TRR 237: Nukleinsäure-Immunität/369799452//Mobile genetische Elemente als Quellen endogener immunstimulatorischer Nukleinsäuren/B19

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