TransIT-Lenti Transfection Reagent

Product No. Quantity Price Add to Cart

TransIT®-Lenti Transfection Reagent

MIR 6603 0.3 ml $120.00

MIR 6604 0.75 ml $300.00

MIR 6600 1.5 ml $475.00

MIR 6605 5 x 1.5 ml $2,066.00

MIR 6606 10 x 1.5 ml $3,800.00

TransduceIT™ Transduction Reagent

MIR 6620 1 ml $25.00

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

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Product Overview

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

High Performance – Provide up to eight-fold higher functional titers
Simple Protocol – No media change required, single harvest
Animal Origin Free – Regulatory friendly

Glossary of Lentivirus Terms Click to expand

Term	Definition
Functional Titer	A measure of the amount of infectious virus particles (e.g. containing envelope, capsid and genome). Functional titer is often reported in Transducing Units per milliliter (TU/ml).
LTR	Long Terminal Repeats are regions of the RNA genome associated with regulation, integration and expression of retroviruses. In a lentiviral transfer plasmid, LTR sequences flank the gene of interest and aid in generating viral dsDNA by reverse transcription and integration into the host genome.
MOI	Multiplicity of Infection refers to the ratio of the number of virus particles to the number of target cells.
Packaging Plasmids	Packaging plasmids include the essential structural (gag), enzyme (pol), packaging (rev) and envelope (VSV-G) components of the lentivirus genome. For 2nd generation packaging plasmids, gag, pol and rev are encoded on a single vector, whereas VSV-G is on a separate vector. For increase safety, 3rd generation vectors separate gag and pol from rev in addition to maintaining VSV-G on a separate vector. Third generation vectors often have additional safety parameters such as fewer wild-type HIV sequences and self-inactivating (SIN) elements.
P24 Protein	P24 is a protein component of the lentivirus capsid. Assays that detect P24 protein are commonly used to quantify lentivirus. However, these assays overestimate functional virus titers because they detect defective virus particles and free P24 protein in addition to competent particles. For lentivirus, 100-1000 defective virus particles are produced for every functional virus particle.
Transdcution	The process by which foreign DNA is introduced to the cell by a virus.
Transduction Reagent	A chemical compound that increases virus aggregation and uptake into a target cell type.
Transfection	Transfection is the process by which molecules such as DNA, RNA, oligonucleotides and protein are delivered to cells by non-viral (e.g. chemical or mechanical) methods. In the context of recombinant virus production, transfection is used to deliver plasmid DNA encoding the essential virus components (packaging plasmids) and the gene of interest (transfer plasmid) to producer cells such as HEK 293T/17.
Transfer Plasmids	The transfer plasmid encodes the gene of interest flanked by long terminal repeat (LTR) sequences, as well as other essential components such as RNA packaging signal (Psi) and rev response element (RRE).
VSVG	The Vesicular Stomatitis Virus G glycoprotein is a broad tropism envelope protein that is frequently used to pseudotype recombinant lentivirus. VSV-G also increases lentivirus stability in downstream purification processes.

Overview of Recombinant Lentivirus ProductionClick to expand

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Figures and Data

High Functional Titers with TransIT®-Lenti Transfection Reagent
High Transduction Efficiency in Neuron Cells with Unconcentrated Lentivirus Using TransIT®-Lenti
Functionality Comparison of CaPO4, Lipofectamine™ 2000 or TransIT®-Lenti Generated Lentivirus.
High Efficiency Transfection with TransIT®-Lenti Transfection Reagent
Lentivirus Production is Scalable

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 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 the MISSION® vectors (pLKO.1-puro-CMV-TurboGFP™ transfer vector and the Lentivirus Packaging Mix powered by MISSION®). 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 withlentivirus 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.

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. HT1080 cells 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 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.

Lentivirus Production is Scalable. Adherent 293T/17 cells were transfected in a 12-well, 6-well or 100 mm plate format using the MISSION® vectors (pLKO.1-puro-CMV-TurboGFP™ transfer vector and the Lentivirus Packaging Mix 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.

Applications

Virus Production CRISPR Genome Editing Protein Production Stem Cell Transfection

Product Resources

Specifications

Storage Conditions:
All Configurations: Store at -20°C

Product Guarantee:
All Configurations: 6 months

Technical Product Literature

DownloadProtocol DownloadQuick Ref Protocol DownloadData Sheet SDSTransIT®-Lenti Transfection Reagent SDSTransduceIT™ Transduction Reagent

Additional Resources

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TransIT®-Lenti Transfection Reagent

Ideal for Recombinant Lentivirus Production