Mirus – TransIT-VirusGEN® SELECT Transfection Reagent and Kits
Large Scale Virus Production for Preclinical and Early Phase Clinical Trials
 |
NEW! Click here to order or request a quote!
|
VirusGEN® SELECT AAV Transfection Kit – For suspension 293 cells
Kit includes:
TransIT-VirusGEN® SELECT Transfection Reagent (30 mL or 150 mL)
VirusGEN® SELECT AAV Complex Formation Solution and Enhancer (1 L or 5 x 1 L)
Click here to order or request a quote.
VirusGEN® SELECT LV Transfection Kit – For suspension or adherent 293 cells
Kit includes:
TransIT-VirusGEN® SELECT Transfection Reagent (30 mL or 150 mL)
VirusGEN® SELECT LV Complex Formation Solution (1 L or 5 x 1 L)
VirusGEN® SELECT LV Enhancer (1 L or 5 x 1 L)
Click here to order or request a quote.
TransIT-VirusGEN® SELECT Transfection Reagent is designed to enhance delivery of packaging and transfer vector DNA to suspension and adherent HEK 293 cell types in order to increase production of recombinant lentivirus and adeno-associated virus (AAV). Key benefits of TransIT-VirusGEN® SELECT Transfection Reagent include:
 |
Performance – Efficient DNA delivery for large-scale production of high-titer viral vectors |
|
Flexibility – Compatible with different virus production platforms and repeat filtration |
|
Animal Origin Free – Fully synthetic transfection reagent formulation |
|
Quality – Tested for performance, appearance, identity, sterility, endotoxin and mycoplasma |
With performance that exceeds PEI and PEIpro® reagents, TransIT-VirusGEN® SELECT offers a simplified, cost-effective workflow, making it an excellent choice for large-scale virus production in preclinical studies and early-phase clinical trials. Titers can be further increased with optimized enhancers included in the VirusGEN® SELECT AAV Transfection Kit and VirusGEN® SELECT LV Transfection Kit. When transitioning to late-phase clinical trials or commercial manufacturing, we recommend TransIT-VirusGEN® GMP.
TransIT-VirusGEN® and the VirusGEN® AAV Transfection Kit Produce High AAV Titers at the 1L Scale.
(A) Expi293F™ cells (Thermofisher) grown in Expi293™ Expression Media (Thermofisher) were used to generate recombinant AAV2 via transient transfection using TransIT-VirusGEN® Transfection reagent (1.5:1 reagent-to-DNA ratio (vol:wt), Mirus Bio LLC), VirusGEN® AAV Transfection Kit (Mirus Bio LLC), PEIpro® (1:1, Polyplus) or FectoVIR®-AAV (1:1, Polyplus) using their recommended protocol. AAV2 was produced by transfecting pAAV-hrGFP, pAAV-RC and pAAV-Helper plasmids (1:1:1 DNA ratio, 2 ?g/ml- 2 mg per flask, Agilent Technologies). Cells were transfected at a cell density of 2 million cells/ml, 1 L in a 2.8 L Thomson shake flask. AAV2 was harvested at 72 hours post-transfection using chemical lysis. Functional titers were determined via transduction of HT-1080 cells and GFP expression was measured 48 hours post-transduction using Guava® easyCyte™ 5HT Flow Cytometer. AAV functional titers were measured from virus dilutions with less than 20% GFP positive cells. Genome copies were determined by ddPCR using primers and a probe targeting the CMV promoter.
TransIT-VirusGEN® Reagent and the VirusGEN® LV Transfection Kit Outperform PEI-based Reagents with Multiple Lentivirus Packaging Systems.
Lentivirus production using TransIT-VirusGEN® Transfection Reagent +/- the VirusGEN® LV Enhancer was compared to PEIpro® (2:1 reagent-to-DNA ratio, Polyplus) and PEI MAX (4:1 reagent-to-DNA ratio, Polysciences) using their recommended protocol. Lentivirus was produced by transfecting Expi293™ cells grown in Expi293™ Expression Media (ThermoFisher) at 4 x 10^6 cells/ml with two different LV vector mixtures (1 ug/ml, 2ug/well) including: (A) 3rd generation vectors pALD-LentiEGFP-A transfer vector and pALD-VSV-G-A, pALD-Rev-A, pALD-GagPol-A packaging vectors (3:0.5:0.5:2 DNA ratio, Aldevron) or (B) 3rd generation vectors pLKO.1-puro-CMV-TurboGFP™ transfer vector (MilliporeSigma) and ViraSafe™ Pantropic Packaging mix (pRSV-Rev, pCMV-VSV-G, pCgpV, CellBio Labs) at a 3:0.5:0.5:2 DNA ratio, The VirusGEN® LV Enhancer was added to the TransIT-VirusGEN® condition at 18 hours post-transfection. Virus containing supernatant was used to transduce 293T/17 cells and GFP expression was measured at 72 hours post post-transduction using Guava® easyCyte™ 5HT Flow Cytometer. Lentivirus functional titers were measured from virus dilutions with less than 20% GFP positive cells. The error bars represent the range of duplicate wells.
Technical Product Literature – Full Protocols
TransIT-VirusGEN® SELECT Reagent Full Protocol (PDF)
VirusGEN® SELECT AAV Transfection Kit Protocol (PDF)
VirusGEN® SELECT LV Transfection Kit Protocol (PDF)
Contact our Technical Team for more information.