• Recipient Instructions: Safety, Storage and Use


    Vigene recommends the following steps for safe handling and storage of your virus. Vigene sends all viral particles on dry ice, with over night next day delivery, where applicable. Vigene ships to international customers are the beginning of the week (Monday or Tuesday, Eastern Time USA) with enough ice to keep viral particles stable for no more than 48 hours at ambient room temperature. If your virus did not arrive on dry ice, please contact Vigene immediately.

    Safety

    Storage

    Working with your Virus

    Safety

    Recombinant viruses are replication deficient and can infect a broad range of mammalian cell types. Please refer to your institutions Occuptation Health and Safety office for guidance on safe handling of viral particles. For general information on Biosafety levels refer to the CDC's section on recognizing biosafety Levels.

    rAAV

    Recombinant AAV constructs produced in the absence of a helper virus, that do not encode for either a potentially tumorigenic gene product or a toxin molecule, can be handled in a Biosafety Level 1(BSL-1) facility; if dealing with biohazardous material then please handle under Biosafety Level 2 (BSL-2) containment.

    Vigene does not stock wild-type AAV.

    Lentivirus

    Lentiviral vector constructs are derived from HIV and are therefore highly efficient vehicles for in vivo gene delivery. Their ability to integrate transgenes into dividing, as well as, non-dividing cells makes them extremely desireable. There are risks associated with working with lentiviral vectors; these must be carefully evaluated.

    The two major risks:

      1. potential generation of replication competent virus;
      2. potential for oncogenesis through insertional mutagenesis.

    NIH guidelines require the maintenance of a BSL-2 or BSL-2 enhanced control facility for work involving lentivirus.

     

    Return to Top

     

    Storage

    Vigene sends Lentiviral and AAV particles on dry ice. Upon receipt the receipient should store these particles immediately at -80C. We recommend aliquoting your virus into desired volumes, snap-freezing in liquid nitrogen or a dry ice/ethanol bath before storing at -80°C for use in future experiments. Please store your virus in the back of the freezer to avoid/reduce temperature variations with opening/closing the freezer door. Avoid repeated freeze-thaw cycles with your virus. Temperature fluctuations and freeze-thaw cycles can reduce viral titer.

     

    Return to Top

     

    Working with your Virus

    Thawing prior to use

    Lentiviral particles: thaw, on ice, immediately before use, keeping on ice at all times during experiment.

    AAV particles: thaw, on ice or at room temperature, immediately before use, keeping on ice at all times during experiment.

    Cell Transduction

    ● AAV particles from both small scale and large scale packaging can be applied directly onto cells for cell transduction
    ● Start cell transduction at an AAV MOI of 104 and 106 GC/cell - when cells are readily transducible. With some cell lines a higher AAV MOI might be needed.
    ● Look for the highest transduction with minimal cell death. With some cell lines, high transduction levels cannot be achieved.

    In vivo: Animal Injections

    ● AAV particles from large scale AAV packaging service can
    be applied directly for in vivo animal injection.
    ● The recommended titer for in vivo animal injection is 1011 GC per mouse or 2X109 GC/g (body weight). For local injections (e.g. into brain) a smaller quantity of AAV viruses can be applied.

    Additional Viral Resources

    Ordering Instructions: Shipping and payment information
    Product Manuals: How to use your virus
    FAQs : Answers to commonly asked questions
    Publications: showcasing Vigene products and services

     

    Return to Top