TransIT®-Lenti transfection reagent from Mirus Bio enables enhanced delivery of the essential vectors required for high-titre lentivirus production in adherent HEK 293T and suspension 293-F cells. Using a simple protocol that requires no media change and a single harvest, the animal-origin free TransIT-Lenti gives greater than two-fold higher functional titers over other transfection reagents.
Benefits of using TransIT-Lenti
• High functional titres
• Simple protocol
• No media change required
• Single harvest
• Animal-origin free
High functional titers with TransIT-Lenti transfection reagent
Adherent 293T/17 cells were transfected in a 6-well plate with pLKO.1-puro-CMV-TurboGFP™ transfer vector and the Lentivirus Packaging Mix Powered by MISSION® Genomics (1:1 ratio, 2 µg/well) with the following reagents: TransIT-Lenti (3:1, vol:wt), Lipofectamine® 2000 (3:1), Lipofectamine® 3000 (3:1:1), 25 kDa PEI (6:1), or CaPO4 precipitation (4 µg pDNA/well). 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. Error bars represent triplicate transfection complexes titered individually. Functional titers were calculated using virus dilutions with less than 20% GFP positive cells.
Comparison of CaPO4, Lipofectamine® 2000 and TransIT-Lenti generated lentivirus
HIV CMVeGFP Virus was produced in HEK 293FT cells using either CaPO4, Lipofectamine® 2000 or TransIT-Lenti Transfection Reagent per the manufacturer's protocol. Lentivirus was collected 48 hours post-transfection and concentrated by prolonged centrifugation at 9,000 x g. HT1080cells were infected with a 1:100 or 1:1000 dilution of each concentrated lentivirus. Images (above) were captured 48 hours post-transduction. Data courtesy of Jeremy Coffin, University of Iowa Viral Vector Core
High efficiency transfection with TransIT-Lenti transfection reagent
Adherent 293T/17 cells were transfected in a 6-well plate format using the pLKO.1-puro-CMV-TurboGFP™ transfer vector and the Lentivirus Packaging Mix Powered by MISSION® Genomics using the TransIT®-Lenti Transfection Reagent (3:1, vol:wt). GFP efficiency was measured at 48 hours post-transfection using guava easyCyte™ 5HT Flow Cytometer. Error bars represent five transfection complexes. Images were captured at 48 hours post-transfection (10X objective) using a Zeiss Axiovert S100 inverted fluorescence microscope. The observed cell rounding and cell-cell fusion is due to high expression of the vesicular stomatitis virus G protein (VSV-G) for pseudotyping the recombinant lentivirus.
TransIT Lentivirus System
The TransIT Lentivirus System from Mirus Bio combines the benefits of TransIT®-Lenti with the Lentivirus Packaging Mix powered by MISSION® Genomics to produce even higher functional titers.
Material available for download
TransIT-Lenti Transfection Reagent protocol
TransIT-Lenti Transfection Reagent quick reference protocol
TransIT-Lenti Transfection Reagent SDS
TransIT Lentivirus System protocol
TransIT Lentivirus System quick reference protocol
TransIT Lentivirus System SDS
TransduceIT™ Reagent product data sheet
Note: product availability depends on country - see product detail page.
|Details||Cat number & supplier||Size||Price|
|TransIT®-Lenti Transfection Reagent MIR 6603 · Mirus||
|TransIT®-Lenti Transfection Reagent MIR 6604 · Mirus||
|TransIT®-Lenti Transfection Reagent MIR 6600 · Mirus||
|TransIT®-Lenti Transfection Reagent MIR 6605 · Mirus||
|5 x 1.5 ml||
5 x 1.5 mlview
|TransIT®-Lenti Transfection Reagent MIR 6606 · Mirus||
|10 x 1.5 ml||
10 x 1.5 mlview