Ideal for Recombinant Adeno-associated Virus and Lentivirus Production
TransIT-VirusGEN® Transfection Reagent is designed to enhance delivery of packaging and transfer vectors to adherent and suspension HEK 293 cell types to increase recombinant adeno-associated virus (AAV) and lentivirus production.
- Reliable – Consistent high virus titer production
- Scalable – Efficient across different formats
- Flexible – Compatible with different recombinant virus and cell culture systems
- Animal Origin Free – High performance with maximum compatibility
- Now with increased specification for the functional assay from 5 x 10^5 TU/ml to 5 x 10^6 TU/ml
Additional Resources
Technical Product Literature
Full Protocols
TransIT-VirusGEN® Full Transfection Protocol (PDF)
Quick Reference Protocols
TransIT-VirusGEN® Quick Ref Protocol (PDF)
SDS
TransIT-VirusGEN® SDS (PDF)
Figures and Data
TransIT-VirusGEN® Outperforms Competitor Reagents in Suspension & Adherent AAV Cell Cultures. Suspension FreeStyle™ 293-F cells grown in FreeStyle™ F17 Medium (A) or adherent 293T/17 cells (B) were transfected with pAAV-hrGFP, pAAV-RC, and pAAV-Helper (1:1:1 ratio, 1.5 µg/ml, Agilent Technologies) with the following reagents: TransIT-VirusGEN® (2:1, vol:wt), Lipofectamine® 2000 (3:1, Thermo Fisher), Lipofectamine® 3000 (3:3:1, Thermo Fisher) or 25 kDa PEI (4:1, Polysciences). 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® Outperforms Competitor Reagents in Suspension and Adherent Lentivirus Cell Cultures. Suspension FreeStyle™ 293-F cells grown in FreeStyle™ F17 Medium (A) or adherent 293T/17 cells (B) were transfected with pLKO.1-puro-CMV-TurboGFP™ transfer vector and the Lentivirus Packaging Mix powered by MISSION® (1:1 ratio, 1 µg/ml) with the following reagents: TransIT-VirusGEN® (3:1, vol:wt), Lipofectamine® 2000 (2:1 (A) or 3:1 (B), Thermo Fisher), Lipofectamine® 3000 (2:2:1 (A) or 3:3:1 (B), Thermo Fisher) or 25 kDa PEI (4:1, Polysciences). The supernatant was harvested, filtered (0.45 µm), and titered using 293T/17 cells. Lentivirus transductions were performed in the presence of 8 µg/ml TransduceIT™ and GFP expression was measured 72 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). Suspension FreeStyle™ 293-F cells grown in FreeStyle™ F17 Medium (A) or adherent 293T/17 cells (B) were transfected with pAAV-hrGFP, pAAV-RC, and pAAV-Helper (1:1:1 ratio, 1.5 total µg/ml, Agilent Technologies) using the TransIT-VirusGEN® Transfection Reagent (2:1 reagent-to-DNA ratio (vol:wt), Mirus) at the indicated volumes perculture 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.
Higher Transfection Efficiency with TransIT-VirusGEN® Over PEI for Lentivirus Generation. HEK293T/17 cells were transfected with lentiviral packaging plasmids and either (a) control RFP transfer plasmid or (b) targeting SPdCas9-RFP transfer plasmids using either TransIT-VirusGEN® Transfection Reagent (3:1 reagent to DNA ratio; vol:wt) or 25 kDa linear PEI (3:1 reagent to DNA ratio; wt:wt). Brightfield and RFP Images were taken 17-22 hours post-transfection.
