Generation of Stable Cell Lines by Retrovirus-Mediated Gene Transfer Techniques
Using a retrovirus vector system to generate stable cell lines has several distinct advantages over using other vector systems:
- A retroviral vector is able to transform its single-stranded RNA (ssRNA) into a double-stranded DNA (dsDNA) molecule and stably integrate this dsDNA into a host genome at the transcription active site.
- All viral structural genes of the retrovirus are removed from the expression vector to prevent revertant recombination, making the retroviral vector a safe and powerful tool for gene transfer.
- Retroviral vector-mediated gene transfer has been widely applied in the development of gene therapy.
- Retroviral vectors are compatible with dividing cells and other primary cells.
- The retroviral vector, which is capable of infecting both mouse and human cells, is based on Moloney murine and leukemia viruses (Mo-MLV).
- The viral genes (e.g., gag, pol, and env) in the retroviral system are replaced with the transgene and expressed in trans in the packaging cells.
Cell Sorting to Select Integrated Cells