Recombinant AAV and lentiviral production: TransIT-VirusGEN

TransIT-VirusGEN® is a high-performance transfection reagent engineered for the production of recombinant AAV and lentiviral vectors. Combining proprietary polymer and lipid technologies, VirusGEN delivers efficient DNA delivery and consistently high viral titres in both adherent and suspension HEK293 cell systems. Designed for flexibility and scalability, it supports applications ranging from early-stage research to process development and large-scale manufacturing.

Benefits of using TransIT-VirusGEN

• Consistent high virus titre production
• Compatible with adherent or suspension HEK293 cells
• Can be scaled efficiently
• Streamlined virus generation workflow
• Compatible with different virus & cell culture systems

TransIT-VirusGEN outperforms competitor reagents in suspension and adherent AAV cell cultures

Figure 1. Suspension (A) or adherent cells (B) were transfected with pAAV-hrGFP, pAAV-RC, and pAAV-Helper (1:1:1 ratio) with the following reagents: TransIT-VirusGEN (2:1, vol:wt), Lipofectamine® 2000 (3:1), Lipofectamine® 3000 (3:3:1) or 25 kDa PEI (4:1). Harvested virus was used to transduce HT1080 cells and GFP expression was measured 48 hours post-transduction using guava easyCyte™ 5HT flow cytometer. Functional titers were measured from virus dilutions with less than 20% GFP positive cells. The error bars represent the standard deviation of triplicate wells.

TransIT-VirusGEN enables broad scalability (AAV)

Figure 2. Suspension (A) or adherent cells (B) were transfected with pAAV-hrGFP, pAAV-RC, and pAAV-Helper (1:1:1 ratio) using the TransIT-VirusGEN transfection reagent (2:1, vol:wt) at the indicated volumes per culture vessel. Harvested virus was used to transduce HT1080 cells and GFP expression was measured 48 hours post-transduction by flow cytometry. Functional titers were measured from virus dilutions with less than 20% GFP positive cells. Error bars represent the standard deviation of triplicate wells, flasks, or dishes.

Enhance AAV Production with VirusGEN® and RevIT™ AAV Enhancer 

For enhanced AAV production, RevIT™ AAV Enhancer can be incorporated into the workflow to further increase viral yields and improve overall process efficiency.  

For large-scale AAV manufacturing, TransIT®-AAViator™ provides an advanced transfection platform designed to support high-yield production workflows 

These technologies provide a flexible and scalable platform for viral vector production, supporting everything from early-stage research through to process development and manufacturing. 

• Fewer reactors, less media, less DNA 
• Fewer production runs, less hands-on time
• Efficient use of time and materials, higher profitability
• Higher titers, significant cost savings

Figure 3. VirusGEN plus RevIT AAV enhancer outperformed polymer-only reagents in both titer and percent full capsids.

Figure 4. Reduction in cost per dose driven by titer increase with VirusGEN transfection platform and RevIT AAV enhancer. *Process conditions: 200L bioreactor run in GMP setting, 3M cells per mL, Reagent : DNA ratios per supplier recommendations RevIT AAV enhancer added post transfection Dose: 5E+14, AAV8.

Material available for download

• TransIT-VirusGEN protocol
• TransIT-VirusGEN quick reference protocol for AAV generation
• TransIT-VirusGEN SDS
• How To Titer Virus reference card

Lipofectamine is a registered trademark of Life Technologies Corporation
Guava and EasyCyte are registered trademarks of Guava Technologies, Inc.