Free Sample

TransIT®-Lenti Transfection Reagent

Ideal for Recombinant Lentivirus Production

To inquire about bulk pricing, please call 888-530-0801
International inquiries please call +1-608-441-2852
View Shipping Information

Choose a Product:

Select a Size:

Catalog number selected: -

-

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.

  • High Performance – Provide higher functional titers 
  • Simple Protocol – No media change required, single harvest
  • Animal Origin Free  Regulatory friendly
 
Glossary of Lentivirus Terms Click to expand
Overview of Recombinant Lentivirus ProductionClick to expand

Figures and Data

Copy Link Enlarge Image
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® 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.

Copy Link Enlarge Image
High Transduction Efficiency in Neuron Cells with Unconcentrated Lentivirus Using TransIT-Lenti

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.

Copy Link Enlarge Image
Functionality Comparison of CaPO4, Lipofectamine 2000 or TransIT-Lenti Generated Lentivirus.

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

Copy Link Enlarge Image
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 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.

Copy Link Enlarge Image
Lentivirus Production is Scalable

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.

Resources

COA Lookup

Download Certificates of Analysis

COO Lookup

Download Certificate of Origin

Testimonials

"Excellent technical support, they are very knowledgeable."

Review published in Select Science

Questions?

Contact Technical Support
8:30AM - 4:00PM CST, M-F
EMAIL - techsupport@mirusbio.com
PHONE - 888.530.0801