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TransIT-Lenti Transfection Reagent – 10 x 1.5 mL

Ideal transfection reagent for recombinant lentivirus production

SKU MIR 6606 Categories ,

$4,761.00

Description

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.

The TransIT® Lentivirus System combines the benefits of TransIT®-Lenti with the 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

 

SKU: MIR 6606

Supporting Data

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.

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Specifications

Storage Conditions
TransIT®-Lenti Transfection Reagent: Store at -20°C
Lentivirus Packaging Mix Powered by MISSION® Genomics: Multiple storage conditions – see individual bottles for specific recommendations
TransIT® Lentivirus System: Store at -20°C
TransduceIT™ Transduction Reagent: Store at -20°C

Product Guarantee
TransIT®-Lenti Transfection Reagent: 1 year
Lentivirus Packaging Mix Powered by MISSION® Genomics: 1 year
TransIT® Lentivirus System:
TransduceIT™ Transduction Reagent: 6 months

Usage Statement
All Configurations: For Research Use Only.

Animal Origin Statement
All Configurations: This product is animal origin free.

 

Technical Product Literature

Full Protocols
TransIT® Lentivirus System Full Protocol (PDF)
TransIT®-Lenti Full Transfection Protocol (PDF)

Quick Reference Protocols
TransIT® Lentivirus System Quick Ref Protocol (PDF)
TransIT®-Lenti Quick Ref Protocol (PDF)

Product Data Sheets
Lentivirus Packaging Mix Product Data Sheet (PDF)
TransduceIT™ Reagent Product Data Sheet (PDF)

SDS
Lentivirus Packaging Mix SDS (PDF)
TransduceIT™ Reagent SDS (PDF)
TransIT® Lentivirus System SDS (PDF)
TransIT®-Lenti SDS (PDF)

FAQs

See the TransIT-Lenti FAQs

Using transfection reagents and enhancers from Mirus has given our platform a competitive advantage. Increasing efficiency and lowering costs for all of our porgrams.

 

Sally Mader, PhD
ZYZ Theraputics