TransIT®-Lenti for High Titer Lentivirus Production

TransIT-Lenti Transfection Reagent is designed to enhance delivery of packaging and transfer vectors to adherent HEK 293T and suspension 293-F cell types to increase recombinant lentivirus production.

  • Provide up to eight-fold higher functional titers
  • Simple protocol - no media change required, single harvest
  • Animal origin free formulation

Glossary of Lentivirus Terms Click to expand
Overview of Recombinant Lentivirus Particles and Infection of Target Cell

Overview of Recombinant Lentivirus Particles and Infection of Target Cell. (1) Producer cells (e.g. 293T) are transfected with 3-4 plasmids encoding the gene of interest, vesicular stomatitis G protein (VSV-G) and essential virus proteins (e.g. gag, pol and rev). (2) Virus is assembled and released into the supernatant through budding with the producer cell plasma membrane resulting in an envelope decorated with VSV-G. The medium containing virus is filtered through a 0.45 µm filter to remove any cells. (3) Target cells are frequently transduced with recombinant lentivirus particles in the presence of a polycation to increase aggregation and uptake. The virus enters the cell and the capsid is uncoated revealing the RNA genome and viral enzymes. The viral RNA is reverse transcribed into DNA which is then integrated into the host genome. (4) Transcription and translation result in the production of the protein encoded by the gene of interest.

High Functional Titers with TransIT-Lenti Transfection Reagent

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® (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.

High Efficiency Transfection with TransIT-Lenti Transfection Reagent

High Efficiency Transfection with TransIT®-Lenti Transfection Reagent. Adherent 293T/17 cells were transfected in a  6-well plate format using the MISSION® vectors (pLKO.1-puro-CMV-TurboGFP™ transfer vector and the Lentivirus Packaging Mix) 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.

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Technical Resources

Mirus' Transfectopedia®: Why use recombinant viruses for nucleic acid delivery
Product Details: Find more information at the following product pages:

DNA Virus Production RNA Virus Production Baculovirus Production
TransIT®-VirusGEN™ Transfection Reagent
TransIT®-mRNA Transfection Kit TransIT®-Insect Transfection Reagent
TransIT®-Lenti Transfection Reagent
TransIT®-LT1 Transfection Reagent
TransIT®-293 Transfection Reagent
TransIT®-2020 Transfection Reagent

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