Lentiviral Transduction

TaKaRa Bio

1) Lentiviral Packaging

Lenti-X Packaging Single Shots

• Extremely simple and consistent one step method for producing high titer generating VSVg pseudotyped lentivirus
• The single tube format consists of individual vials containing lyophilized Xfect Transfection Reagent with Lenti-X lentiviral packaging plasmids

 

Lenti-X HTX Packaging System

• Fourth generation of packaging systems for generating VSV-g pseudotyped lentivirus

 

Lenti-X HTX Ecotropic Packaging System

• Fourth generation of packaging systems for generating ecotropic pseudotyped lentivirus

 

Lenti-X HTX Packaging System (Integrase Deficient)

• Packaging system with mutant integrase that minimizes the proviral integration

 

Lenti-X 293T Cell Line

• Special 293T clone selected for high titer lentivirus production.

 

2) Lentiviral Particles

TaKaRa Bio with cooperation with Vectalys produced pecial lentiviral particles with 100% transduction of many primary cells (including neurons, endothelial cells and stem cells) without cytotoxic effects. The particles are in a functional titer what is not just the total particle number, it is a measurement of the total number of virions capable of integrating into cells. The production in a serum-free medium greatly improves the quality of the lentivirus as a vector for gene delivery.

Transduction efficiency of a Vectalys GFP-expressing lentiviral vector in IMR90 cells. Even with a low MOI of 5, >95% of cells are transduced using this high-titer lentivirus. Very high expression can be achieved by increasing the MOI.

 

Viral particles for Tet-Inducible Gene Expression

• Ready made lentiviral particles to introduce constitutive expression of the Tet-On 3G transactivator in the cells
•  In the presence of doxycycline the Tet-On 3G transactivator binds and activates transcription of the P_TRE3G promoter

 

Viral particles for subcellular localization studies

• Constitutive Lentiviral Reporter Vectors are prepackaged lentiviral particles that deliver fluorescent proteins targeted to specific organelles or subcellular structures
•  These particles can be used to create reporter cell lines for the study of cytoskeletal and organelle structure and function in living cells or protein localization studies

 

iPS Cell Production Systems

• Polycistronic recombinant lentiviral particles for generating iPS cells
• These particles encode the Yamanaka factors which transcription generated iPS cells

 

3) Lentiviral Transduction Tools

Identification of integration sites

• Enables simple determination of lentivirus integration sites using GenomeWalker technology
• Compatible with most commercialy available lentivirus vectors

 

Lenti-X Maxi Purification Kit

• Produces high yields of purified virus from crude supernatants and concentrates the virus up to 10 fold
• It uses simple and gentle gravity flow protocol for more efficient and consistent lentiviral transduction and healthier target cells
• It also efficiently removes transduction inhibitors and cytotoxic contaminants

 

Lentivirus Titration

• Lenti-X GoStix Instant Titration test that get a yes or no in only ten minutesC
• Can determine when to harvest lentiviral supernatants only from 20 µl of the lentiviral supernatant
• The other titration kits offered by TaKaRa Bio are used based on methods like ELISA, qRT-PCR and qPCR

 

Lenti-X Accelerator

• Magnetic bead based technology designed to accelerate lentiviral transduction experiments
• The workflow is complete in only 30 minutes and avoid using Polybrene
• Ideal for transducing polybrene-sensitive cells

 

Lenti-X Concentrator

• Simple mix and spin protocol without need of ultracentrifugation
• It provides a fast, simple and efficient method for concentrating any lentiviral stock

 

Ecotropic Receptor Booster

• Used to temporarily elevate the density of the ecotropic receptor protein mCAT-1 on the surface of any target cell
• It allows to efficiently transduce human cells with ecotropic retrovirus or lentivirus that otherwise could only infect rodent cells
• It also increase the infection efficiency of any type of cells that are resistant to viral infection.

 

4) Lentiviral Vector Systems
TaKaRa Bio offers a variety of highly optimized lentiviral vectors for many applications.

Lentiviral Vector Systems for Constitutive cDNA Expression

• Express the gene of interest from a strong CMV promoter
• Select for lentiviral integration using antibiotic selection

 

Bicistronic Lentiviral Vector Systems

• Coexpress the gene of interest and an antibiotic or fluorescent selectable marker from the same transcript
• Using IRES-containing lentiviral vector systems

 

Lentiviral Vector Systems with an EF-1 Alpha Promoter

• Express the gene of interest from an EF-1 alpha promoter in cell types
• CMV promoter can be silenced over time such as hematopoietic or stem cells

 

Lentiviral All-In-One Vector Systems for Inducible Expression of cDNA

• Tightly controlled, inducible expression clones of your gene of interest with only a single vector

 

Lentiviral Vector Systems to Control Protein Stability

• Clone the gene downstream of a ProteoTuner destabilization domain (DD)
• Control the stability of your protein with Shield1 a small molecule ligand which protects the DD fusion protein from being degraded

 

Lentiviral Vectors for Fluorescent Protein Expression

• Clone the gene in frame with one of many fluorescent proteins (FP)
• Monitor the expression and subcellular localization of the protein of interest

 

Promoter Studies with Lentiviral Vectors and Secreted Luciferase

• Monitor activity of the favorite promoter using a reporter system that expresses bright fluorescent proteins or secreted luciferase

 

Promoter Studies with Lentiviral Vectors and Fluorescent Proteins

• Monitor activity of the favorite promoter using a reporter system that expresses a cyan, green or red fluorescent protein

 

Documents and resources

For more information visit producer's website.