Retroviruses are enveloped viruses that are found in fish, avian and mammals. The diameter of a retroviral virion is approximately 80 – 100 nm. The outmost layer of the virus is a lipid bilayer derived from the host cell membrane. It surrounds the spherical capsid in which two identical single stranded RNA strands are located.

They are positive-sense and approximately 7 – 11 kB long [5]. Generally the retroviral life cycle can be divided in two distinct phases: infection and replication. In the following sections we will focus on the Moloney Murine Leukemia Virus (MuLV), which is the origin of the viral vector we are using for our project.

Viruses are classified as ecotropic, polytropic or amphotropic depending on the number of different host cells they are able to infect [1]. The Moloney Murine Leukemia Virus (MuLV) is an ecotropic gammaretrovirus; it only infects rodent cells (mice and rat). This tropism of the virus is granted by its high specificity towards the mCAT-1 receptor [7]. Once attached to the receptor the MuLV enter the target cell and the single stranded viral RNA is transferred into the cytoplasm.

MuLV is a simple retrovirus naturally encoding only three genes called gag, pol and env. These so called trans-gene elements are flanked by long terminal repeats (LTR). The 5’ LTR harbors a promoter region, which initiates transcription of the provirus, whereas the 3’ LTR is needed for polyadenylation of the proviral mRNA. The gag (group specific antigen) genes are precursor polyproteins that form the major components building up the core particle, RNA binding proteins and the nucleoprotein core particle. The pol genes encode for the reverse transcriptase, RNase H and the Integrase. It is mandatory for proper processing of the viral RNA genome. The env gene codes for the envelope protein. This protein is responsible for the viral tropism and is located in the viral lipid bilayer which is derived from the host cell membrane. The retroviral psi-packaging sequence is a cis-acting RNA element which allows the transcribed viral RNA to be incorporated into the assembly of the new virus [3]. After proper packaging the virus is leaving the cell and can start the next round of infection.

Retroviral vectors are used to deliver variable gene cargos into cells, a process termed transduction. To ensure safe handling the cis- and trans-acting gene elements are separated from each other. Retrovirus vectors can be made in the absence of helper virus by using so called packaging cell lines. These cell lines carry the trans-acting gene-elements, gag, pol and env, which are needed to produce a functional viral particle. Besides safety issues this approach also has the advantage that a larger gene cargo (~8 kb) can be integrated into the vector [4]. Our MuLV vector needs the Phoenix eco cell line in order to replicate.