Silencing Cargo Gene Expression In Lentiviral Vector And AAV Production To Improve Vector Yield And Purity

By Qian Liu, Sherin Parokkaran Johny, Matthew Tridgett, Mohine Alam, Fran Bennett, Siobhan Clerkin, Michal Fleszar, Meenakshi Raghunath, Maria Patricio, Michael White, Weiheng Su, Vaughan Leydon

Lentiviral and adeno‑associated viral vectors are central to modern cell and gene therapies, yet manufacturing challenges persist when therapeutic cargo genes are highly expressed or cytotoxic. Excessive cargo expression during production can compromise cell viability, generate unwanted byproducts, and place added strain on downstream processing, ultimately limiting yield and process robustness.

One emerging strategy addresses these issues by suppressing cargo gene expression during vector manufacturing while preserving functional vector output. By decoupling vector RNA production from cargo gene expression, targeted silencing approaches allow production cells to remain viable and productive, even when handling difficult or toxic transgenes. The result is improved cell health, reduced host‑cell–derived impurities, and higher recoverable titres.

Implemented across both transient and stable production systems, cargo gene silencing demonstrates measurable gains in productivity and manufacturability for LVV and AAV platforms. These approaches offer a practical path to de‑risking vector production, enabling more consistent scale‑up and supporting the development of increasingly complex gene therapy programs.