• Lentiviral Transduction System


    Vigene Biosciences offers ultra high titer lentivirus production service. Accelerate your discoveries, let Vigene provide you the viral tools you need. Vigene Biosciences has developed the most robust and professional AAV packaging service, ranging from small crude scale to the custom large purified scale. Provides high-quality and high-titer viral services. Receive ready-to-transduce, ultra high titer and high purity lentivirus in as little as 2 weeks.

    High Titer Lentivirus Production

    Lentivirus packaging and purification

    Lentivirus custom cloning

    Custom Lentivirus cloning into any Vigene lentivector

    Lentivirus packaging: different levels of titer and flexible volumes

    ● High titer: 109 IFU/mL option with fast turn around, usually 1-3 weeks

    ● Ready-to-package lentiviral ORFs

    ● Custom cloning: with different promoters, shRNA, TALEN and CRISPR.

    Important: Insert Size limitation and Lentivirus production overview

    Lentivirus

    The lentivirus is enveloped, spherical to pleomorphic in shape, measuring 80-100 nm in diameter. The mature capsid contains 1572 capsid proteins.

    Lentiviruses are a subset of retroviruses. They are some of the most common and useful viruses used throughout research. They have the ability to transduce both dividing and non-dividing cells without significant immune responses. Integrating stably into the host genome they enable long term transgene expression.

    There are some safety considerations to evaluate when working with lentivirus. The Lentivirus is based on HIV-1, which may require additional lab biosafety procedures. Additionally, this virus integrates randomly into the host genome whereby it has the potential for insertional mutagensis. The 2nd and 3rd generation lentiviral systems provide increased researcher safety.

    Lentivirus has broad cell spectrum: infecting most cells, dividing & non-dividing. It is useful for both easy-to-transfect & hard-to-transfect cells. Offering convenience, with minimal need for optimization.

    Lentiviral Services

    Lentivirus Cloning Service

    ● Lentivirus Human cDNA ORF Cloning.

    ● Lentivirus shRNA Cloning.

    ● FLEX-ON of Cre dependent inducible expression.

    Explore lentivirus cloning options.

    Lentivirus Packaging Service

    ● Small Scale Crude Lentiviral Packaging Service.

    ● Large Scale custom Lentiviral Packaging Service.

    ● Lentivirus delivery of Cas9 and sgRNA.

    Explore lentivirus packaging options.

    Related Lentivirus Products

    Control Viruses

    ● Fluorescent proteins: GFP, RFP; Cre recombinase; Tissue-specific promoters.

    Mammalian Expression Vectors.

    ● Shuttling Vectors; shRNA Vectors

    Additional Viral Resources

    Ordering Instructions: Shipping and payment information.

    Product Manuals: How to use your virus.

    FAQs: Answers to commonly asked AAV questions.

    Publications: showcasing Vigene products and services.

    Recipient instructions: information on viral safety and storage.

     

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    Lentiviral Production Overview

    Limitations on insert size for lentivirus packaging and titer

    The titer of lentivirus is very sensitive to the size of the viral genome. Vigene's offered titer is based on the viral genome: the region between the 5’and 3’LTR should be less than 5.1kb, with the gene of interest less than 1.5kb. For every kb increases of the viral genome, the titer will be decreased by ~10 times (drop of 1 log in titer for every 2Kb of insert). Typically, Vigene lentivectors can accommodate a fragment of up to 6kb. Reason for the limitation? There is only so much room within the virus capsid, thereby the integration efficiency goes down, slowing transcription due to the large size. For more inforamtion see article Systematic Determination of the Packaging Limit of Lentiviral Vectors by Mukesh Kumar, Brian Keller, Ndeye Makalou, Richard E. Sutton, Human Gene Therapy. 2001, 12(15): 1893-1905.

    Which lentivirus generation?

    Lentiviral vectors have evolved to achieve higher efficiencies and biosafety. They are composed from 3 components: (1) genomic RNA, (2) internal structural and enzymatic properties, and (3) the glycoprotein envelope.
    1st Generation: composed of all HIV genes minus the envelope protein;
    2nd Generation: Consists of 5 out of the 9 genes deleted, what reamins is the gag/pol and the tat/rev regions; responsible for structural and enzymatic components as well as transcriptional and post-transcriptional genes, respectively.
    3rd Generation: favoured for its clinical applications and considered the safest. Containing only gag, pol, and rev genes and utilizes the chimeric 5’ LTR, which ensures transcription in absence of Tat.

     

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