What is an amphotropic virus?
What is an amphotropic virus?
[ ăm′fə-trŏp′ĭk, -trō′pĭk ] n. An oncornavirus that does not produce disease in its natural host, but does replicate in tissue culture cells of the host species and in cells from other species.
What are Phoenix cells?
Phoenix is a second-generation retrovirus producer cell line for the generation of helper-free ecotropic and amphotropic retroviruses. This cell line is capable of carrying episomes for long-term stable production of retrovirus (LZRS).
What is a retroviral plasmid?
We produce retroviruses by transfecting multiple plasmids that between them provide all of the necessary components for vector particle production. However, the genome plasmid does not encode all, or even any, viral genes. Thus the infected cells make the transgene, eg GFP, but no viral genes.
What is retroviral transduction?
Retroviruses have the ability to transform their single-stranded RNA genome into a double-stranded DNA molecule that stably integrates into the genome of dividing target cells. Retroviral transduction has been widely used for cancer and stem cell research.
Can I freeze retrovirus?
A: VSVG-pseudotyped retrovirus can be frozen, but it will degrade even at -80ºC. On average, each freeze/thaw cycle will result in a 2-4 fold decrease in titer. Amphotropic and ecotropic viruses are not able to handle a freeze/thaw cycle and should be used fresh.
What are Plat A cells?
The Platinum-A (Plat-A) Cell Line, a potent retrovirus packaging cell line based on the 293T cell line, was generated using novel packaging constructs with an EF1α promoter to ensure longer stability and high-yield retroviral structure protein expression (gag, pol, amphotropic env).
What are hek293t cells?
293T (or HEK 293T) is a human cell line, derived from the HEK 293 cell line, that expresses a mutant version of the SV40 large T antigen. It is very commonly used in biology for protein expression and production of recombinant retroviruses.
What is the disadvantages of retrovirus?
Disadvantages and risks of using the retrovirus as a viral vector in gene therapy include low transduction efficiency, replication competence, insert size, integration, inactivation by complement cascade, and the requirement of cell division for transduction.
How do you freeze a stock virus?
Keep the virus stock on ice. Sonicate twice for 30 sec in ice water, with a 30 sec rest on ice between, and divide it into 0.5- to 2-ml aliquots. Store the aliquots indefinitely at −80°C.