Viral vector platform

Lentivirus packaging

Lentiviral vectors integrate into the host genome, which makes them the default choice when a transgene has to be expressed reliably over many passages. FuBio packages lentivirus for overexpression, RNA interference and CRISPR delivery, and supplies catalog constructs for the most common reporter and selection combinations.

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What Are Lentiviral Vectors?

Lentiviruses are a genus of retroviruses. The particle is enveloped and roughly spherical, about 80 to 100 nm across, and carries two copies of a positive-sense single-stranded RNA genome. HIV-1 is the best characterized member and the backbone of most lentiviral vector systems.

Lentiviral vectors are engineered to be replication-deficient and self-inactivating. The viral genes are split across separate packaging plasmids, and the transfer vector keeps only the elements needed for packaging, reverse transcription and integration. Because the vector integrates into the host genome, expression is stable across passages, and unlike gamma-retroviral vectors lentivirus also transduces non-dividing cells.

Schematic of a lentiviral vector particle: a VSV-G-pseudotyped envelope surrounding a conical capsid that contains two copies of the RNA genome, with the long terminal repeats, the packaging signal and the expression cassette labeled.

Lentiviral Vector Packaging and Production

FuBio produces lentiviral vectors using a third-generation packaging system in HEK293T cells. Essential viral components are distributed across separate plasmids to support vector production while minimizing the risk of generating replication-competent lentivirus (RCL).

  • Transfer Plasmid

    Carries the transgene expression cassette, flanked by lentiviral long terminal repeats (LTRs), along with the packaging signal and other cis-acting elements required for vector production and transduction. The self-inactivating (SIN) design includes a deletion in the U3 region of the 3′ LTR, reducing LTR-driven transcription after integration.

  • Packaging Plasmid

    Provides the Gag structural proteins and Pol enzymes required for lentiviral particle assembly, reverse transcription, and integration.

  • Regulatory Plasmid (Rev)

    Expresses the Rev protein, which facilitates the nuclear export of vector RNA containing the Rev response element (RRE), supporting efficient lentiviral vector production.

  • Envelope Plasmid

    Provides the envelope glycoprotein that influences vector tropism and cell entry. VSV-G is commonly used because of its broad host range and stability during vector concentration. Alternative envelope proteins may be available upon request.

Second- and Third-Generation Lentiviral Packaging Systems

Lentiviral vectors are commonly produced using second- or third-generation packaging systems, which differ in plasmid composition, regulatory elements, and transfer vector compatibility.

Comparison of the second-generation (three-plasmid) and third-generation (four-plasmid) lentiviral packaging systems, showing the vector, packaging, envelope and regulatory plasmid constructs with LTR, psi/RRE, promoter and transgene elements.
Figure. Plasmid Composition of Second- and Third-Generation Lentiviral Packaging Systems

Second-generation packaging systems typically provide Gag/Pol, Tat, and Rev functions through a packaging plasmid, together with an envelope plasmid. Third-generation systems separate Rev from Gag/Pol and generally omit Tat, using transfer vectors designed for Tat-independent transcription.

Second-generation packaging systems are compatible with many second- and third-generation transfer vectors, whereas third-generation packaging systems generally require Tat-independent transfer vectors. Actual compatibility depends on the specific vector and packaging plasmid designs.

Lentiviral Packaging Service Advantages

  • Up to 1 × 10⁸
    TU/mL

    Our lentiviral packaging platform has demonstrated a functional titer of 1 × 10⁸ TU/mL. Actual titers may vary depending on the transfer vector, transgene, and other project-specific factors.

  • 15-25
    Business Days

    Our standard turnaround time is 15–25 business days, depending on project requirements and service scope.

  • 2
    Service Options

    Choose from standalone lentiviral packaging using customer-supplied transfer vectors or an integrated service that includes vector construction and packaging.

  • Expert
    Technical Support

    Our team provides technical guidance on vector compatibility, project specifications, and service selection to support your research needs.

Applications of Lentiviral Vectors

  1. 1

    Stable Cell Line Development

    Generating stable pools and clones that keep expressing the transgene across many passages, with early-passage material banked for future work.

  2. 2

    Gene Overexpression and Knockdown

    Delivering cDNA, shRNA and miRNA constructs for gain- and loss-of-function studies in dividing and non-dividing cells.

  3. 3

    CRISPR Delivery and Screening

    Delivering Cas9 and guide RNAs for knockout, activation and interference studies, including pooled library screening.

  4. 4

    Reporter and Biosensor Cell Lines

    Building reporter lines for pathway, promoter and drug-response studies, including inducible and selection-coupled designs.

  5. 5

    In Vivo Delivery in Preclinical Models

    Local and systemic delivery in animal models where long-term expression is needed, with envelope and promoter matched to the target tissue.

Lentiviral Vector Features

Feature Lentivirus
Gene Expression Long-term, stable (integrating)
Immunogenicity Low
Payload Capacity ~8 kb between the LTRs
Tropism (Targeting ability) Broad – VSV-G pseudotyping; envelope can be changed
Best used for Stable lines, primary and non-dividing cells

Production and quality control

  • Vectors are produced by transient co-transfection of HEK293T cells with the transfer, packaging and envelope plasmids; supernatant is harvested 48 to 72 hours after transfection.
  • Crude supernatant is clarified, concentrated and buffer-exchanged before release; no serum-derived stabilizer is added unless the product sheet states otherwise.
  • Functional titer is reported in TU/mL (transducing units per milliliter), measured on a permissive cell line; sterility, mycoplasma and endotoxin are tested on every lot.
  • Replication-competent lentivirus (RCL) testing can be arranged for lots destined for in vivo work — request it when you ask for a quote.

Our Service Options

  • Standalone Lentiviral Packaging

    For researchers with existing transfer vectors, we provide lentiviral packaging services using customer-supplied constructs, subject to compatibility review.

  • Vector Construction & Lentiviral Packaging

    For projects requiring additional support, our integrated service covers vector construction and lentiviral packaging, offering a streamlined solution from vector design requirements to lentiviral vector production.

Service Specifications

Specification Information
Vector Type HIV-1-based lentiviral vector, third-generation packaging system
Producer Cells HEK293T
Envelope VSV-G pseudotype by default; alternative envelopes on request
Vector Titer Up to 1 × 10⁸ TU/mL (functional titer)
Volume 200 μL
Quality Control Functional titer by transduction assay; sterility, mycoplasma and endotoxin on every lot; RCL testing on request
Turnaround Time 15-25 business days
Application In vitro / In vivo Research

Start Your Lentiviral Packaging Project

Whether you need packaging for an existing transfer vector or a comprehensive vector construction and packaging solution, our team is ready to help you identify the right approach for your research.

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Experimental validation

Fluorescent reporter constructs allow lentiviral transduction to be confirmed directly by imaging. The images below show GFP reporter expression after transduction with FuBio lentiviral preparations across adherent and suspension lines: the hepatoma line HepG2, the colon carcinoma line SW480, the diffuse large B-cell lymphoma line OCI-LY8 and the human NK cell line NK-92.

Fluorescence microscopy image of GFP expression in HepG2 cells after lentiviral transduction
Figure 1 · GFP reporter expression in HepG2 cells after lentiviral transduction.
Fluorescence microscopy image of GFP expression in SW480 cells after lentiviral transduction
Figure 2 · GFP reporter expression in SW480 cells after lentiviral transduction.
Fluorescence microscopy image of GFP expression in OCI-LY8 cells after lentiviral transduction
Figure 3 · GFP reporter expression in OCI-LY8 cells after lentiviral transduction.
Fluorescence microscopy image of GFP expression in NK-92 cells after lentiviral transduction
Figure 4 · GFP reporter expression in NK-92 cells after lentiviral transduction.

Resources

User instructions

A condensed bench protocol for transducing cultured mammalian cells with FuBio lentiviral preparations. The full version ships with each order and the current PDF can be requested through the quote form.

  1. 1

    Plan the transduction

    Choose a starting MOI of 1 to 10 for easy-to-transduce lines (HEK293T, HeLa) and 10 to 50 for primary or suspension cells; run a small MOI series the first time. Plate cells so they are 50 to 70 percent confluent on the day of transduction, and include an untransduced control plus, if you will select, a kill-curve plate.

  2. 2

    Thaw and handle the virus

    Thaw a vial quickly in a 37 degree water bath or on ice for larger volumes, mix gently — never vortex — and keep aliquots on ice while working. Avoid more than one freeze-thaw cycle: each one costs roughly an order of magnitude in functional titer.

  3. 3

    Transduce

    Replace the culture medium with fresh complete medium containing the diluted virus. Add polybrene at 4 to 8 micrograms per milliliter only for cell types you have confirmed tolerate it; spinoculation at 800 g for 60 minutes can substitute where polybrene is toxic. Incubate 6 to 16 hours, then replace with fresh medium if the vector or cell type is sensitive.

  4. 4

    Select or enrich

    Start antibiotic selection 48 to 72 hours after transduction at the concentration established by your kill curve (puromycin is commonly 1 to 2 micrograms per milliliter for common lines, but test yours). For fluorescent reporters, enrich by FACS or keep the whole population and record the positive fraction.

  5. 5

    Confirm and expand

    Verify expression by the reporter, flow cytometry, Western blot or qPCR before relying on the line. Expand under selection at the minimum effective pressure, and bank an early passage.

Bench guidance only: optimize multiplicity of infection, selection pressure and timing for your own cell type, and carry out all lentiviral work under the biosafety conditions approved by your institution.

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