To inquire about bulk pricing, please call 888-530-0801 International inquiries please call +1-608-441-2852 Return Policy View Shipping Information
* Additional freight charges may apply for shipments heavier than 10 lbs. Please contact customer service to verify shipping charges: 888-530-0801.
Choose a Product:
Select a Size:
Catalog number selected: -
-
The selected product requires a quote request
Complete the short form below or contact customer service at customerservice@mirusbio.com or +1-608-441-2852 to request a quote for this product.
Complete the short form below or contact customer service at customerservice@mirusbio.com or +1-608-441-2852 to request information about this product.
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. NEW! The TransIT® Lentivirus System combines the benefits of TransIT®-Lenti with the NEW! Lentivirus Packaging Mix Powered by MISSION® Genomics to produce even higher functional titers.
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. 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.
High Transduction Efficiency with Unconcentrated Lentivirus Using TransIT®-Lenti. (A) Lentivirus was produced with the TransIT®-Lenti Transfection Reagent (3:1, vol:wt) or Lipofectamine® 2000 using pLKO.1-puro-CMV-TurboGFP™ transfer vector and the Lentivirus Packaging Mix Powered by MISSION® Genomics). The supernatant was harvested, filtered (0.45 µm), and frozen. Lentivirus transductions were performed 5 days post-plating with iCell® Motor Neurons (Cellular Dynamics International). For bothTransIT-Lenti and Lipofectamine® 2000, one microliter of unconcentrated supernatant was added per well of a 96-well plate. GFP efficiency was measured 72 hours post-transduction using guava easyCyte™ 5HT Flow Cytometer. Error bars represent the SEM of duplicate wells. (B) iCell® Motor Neurons were plated in Ibidy 35mm dishes and transduced with lentivirus produced using the TransIT®-Lenti Transfection Reagent and MISSION® vectors. Images were captured at 72 hours post-transduction with a Zeiss Axiovert S100 inverted fluorescence microscope using a 63X objective under oil.
Comparison of CaPO4, Lipofectamine® 2000 or 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.
Lentivirus Production is Scalable. Adherent 293T/17 cells were transfected in a 12-well, 6-well or 100 mm plate format using the pLKO.1-puro-CMV-TurboGFP™ transfer vector and the Lentivirus Packaging Mix Powered by MISSION® Genomics at a 1:1 ratio, and the TransIT®-Lenti Transfection Reagent (3:1, vol:wt). 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.
TransIT® Lentivirus System Full ProtocolTransIT®-Lenti Full Transfection Protocol
TransIT® Lentivirus System Quick Ref ProtocolTransIT®-Lenti Quick Ref Protocol
Lentivirus Packaging Mix Product Data SheetTransduceIT™ Reagent Product Data Sheet
Lentivirus Packaging Mix SDSTransduceIT™ Reagent SDSTransIT® Lentivirus System SDSTransIT®-Lenti SDS
TransIT-Lenti Transfection Reagent FAQ
Download Certificates of Analysis
For successful COA look up, please use the following tips to locate the appropriate lot number for the respective product categories. If your COA search is not successful using these tips OR you purchased Mirus products before 2011, please request a COA by emailing techsupport@mirusbio.com.
Download Certificate of Origin