What are phoenix cells?

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 amphotropic retrovirus?

Platinum Retroviral Expression System, Amphotropic Retroviral vectors are useful for delivering genes of interest into a target cells where integration into the genome is desired. However, traditional retroviral expression technologies result in low viral titers, making gene expression studies challenging.

How do you freeze retrovirus?

Adeno-Associated Viruses (AAV) Lentiviruses. Retroviruses….DO:

1. Do aliquot and freeze at -80C for long term storage if the virus is not used within ~1 week.
2. Thaw on ice just before use.
3. Use virus within ~ 6 months of storage at -80C unless the titer is much higher than needed.

How do you make a retrovirus?

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.

Can you 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.

How long can you store lentivirus at?

Storage of Lentivirus Virus can be stored at 4°C for a short time (less than a week) before using after reception.

How long is retrovirus genome?

Retroviral genomes are non-segmented, of positive polarity, and typically between 7 to 12 kb long in size.

What is the benefit of using a retrovirus as a vector in gene therapy?

The most important advantage that retroviral vectors offer is their ability to transform their single stranded RNA genome into a double stranded DNA molecule that stably integrates into the target cell genome. This means that retroviral vectors can be used to permanently modify the host cell nuclear genome.

What kind of retrovirus is the Phoenix system?

Phoenix is a second-generation retrovirus producer lines for the generation of helper free ecotropic and amphotropic retroviruses. The lines are based on the 293T cell line (a human embryonic kidney line transformed with adenovirus E1a and carrying a temperature sensitive T antigen co-selected with neomycin).

How to test Phoenix ampho for helper virus?

Phoenix-Ampho have been extensively tested for helper virus production and established as being helper-virus free. Check all containers for leakage or breakage. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.

How is the Phoenix ampho kidney cell line unique?

The unique feature of this cell line is that it is highly transfectable with either calcium phosphate mediated transfection or lipid-based transfection protocols– up to 50% or higher of cells can be transiently transfected. This cell line is capable of carrying episomes for long-term stable production of retrovirus (LZRS).

Which is the second generation retrovirus cell line?

For example, Phoenix, a second generation γ-retrovirus packaging cell line developed by Garry Nolan at Stanford contains Gag-Pol and either an ecotropic envelope, Phoenix-ECO, (for infection of mouse and rat cells), or an amphotropic envelope, Phoenix-AMPHO (for the infection of mammalian cells.)