Most academic institutions classify lentiviral vector production and transduction as BSL-2 work. This applies when you use replication-incompetent, self-inactivating designs with split packaging components. Always consult your institutional biosafety committee before you begin, as local regulations and institutional policies vary.
Cloning & Expression Systems
Lentiviral Expression Systems
Lentiviral expression systems for stable transgene delivery in dividing and non-dividing cells
A lentiviral expression system gives you a direct path from plasmid to a stable, transduced cell line, in both dividing and non-dividing mammalian cells. Packaging mixes come in 2nd- and 3rd-generation configurations. Both split the packaging functions across two or three separate plasmids. This lowers your risk of recombination into replication-competent lentivirus.
Individual packaging plasmids, such as psPAX2 and pMD2.G, are available as catalogue items for custom packaging setups. Ready-to-use packaging mixes that bundle a DNA transfection reagent are also available, if you prefer an all-in-one format.
Cumate-inducible lentivectors use a CuO operator paired with the CymR repressor. They give you dose-controlled stable expression when a constitutive promoter is not suitable.
To generate lentiviral particles for stable transduction, pair a transfer vector with a packaging mix that matches the generation your biosafety protocol allows. Lentiviral vectors integrate semi-randomly into your host genome. Plan for selection cassettes and for downstream validation of integration site and copy number. Use suitable controls to validate and monitor your packaging and transduction process.
Choose your packaging generation and format.
1. Biosafety requirement. If your institution requires 3rd-generation split-packaging, choose a three-plasmid system. This separates the gag-pol, rev, and envelope functions. If 2nd-generation is approved, a two-plasmid packaging mix is the simpler entry point for your setup.
2. Reagent included or separate. Some packaging kits bundle a compatible DNA transfection reagent. Standalone mixes assume you already have a lipofection agent in-house.
3. Inducible expression. When you need tunable transgene dosing, choose cumate-inducible lentivectors using the CuO-CymR system. They let you titrate expression with cumate, instead of running a constitutive promoter at full output from day one.
4. Pseudotyping or library packaging. Pseudotyping with alternative envelopes is available at quoted pricing for your application. Large-scale library packaging services are also available through catalogue service entries.
Parent category: Cloning & Expression Systems
Applications
Stable transgene delivery in non-dividing cells
HIV-1-derived vectors transduce post-mitotic neurons, macrophages and dendritic cells, the preferred route into differentiated cells.
Primary cell line engineering
Hard-to-transfect T cells, haematopoietic stem cells and iPSC-derived populations transduce at low MOI, generating stable reporter or overexpression lines.
Inducible expression studies
Cumate-inducible lentivectors give dose control for toxicity, pathway and rescue work where constitutive overexpression masks a phenotype.
Tropism engineering by pseudotyping
Swapping the envelope for an alternative pseudotype redirects particles toward hard-to-transduce cell types, broadening or restricting tropism.
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Control Plasmids and Premade Viral Particles
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Constitutive cDNA Cloning and Expression Vectors
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Packaging Plasmids and Kits
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Lentivirus Titration and Quantification
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Transduction Reagents
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Lentivirus Purification, Concentration and Preservation
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Promoter Evaluation Kits
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Cumate-Inducible Expression System
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Tetracycline-Inducible Expression System
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Frequently asked questions
Second-generation systems use two packaging plasmids. One encodes gag-pol and rev; the other carries the envelope. Third-generation systems split gag-pol, rev, and envelope across three separate plasmids. This reduces the risk of recombination and of generating replication-competent lentivirus. Biosafety committees often prefer third-generation systems for this extra safety margin.
The most common methods are qPCR-based titration and p24 antigen ELISA. qPCR measures viral RNA or integrated proviral copies. The ELISA measures capsid protein. To find the functional titre, transduce a known number of cells at serial dilutions. Then measure the percentage of transduced cells by flow cytometry or drug selection.
Standard lentiviral transfer vectors accommodate inserts up to approximately 8 kb between the LTRs. This includes the promoter, coding sequence, and selection cassette. Cargo larger than 8 kb shows a sharp drop in titre. For very large transgenes, PiggyBac or adenoviral vectors are more appropriate.
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