Stable Cell Line Generation Lentivirus

This protocol can be used to generate stable cell lines expressing a gene of interest from an integrated lentiviral vector.

Stable cell line generation lentivirus. The authors describe a new lentiviral vector packaging cell line lentipro26 using a mutated less active viral protease to overcome potential cytotoxic limitations. That is many lentiviral genomes. Thus we believe that this study demonstrates that transduction of packaging lines by csin. Lentiviral vectors offer stable integration of your gene of inte.

You may try ecopack 293 to produce the first virus and then use it to infect pt67 to produce the virus you will use for stable cell line generation. The advantage of using the 3rd generation lentivirus are that are very safe and they are replication incompetent. All 2nd generation lentiviral transfer plasmids must be used with a 2nd generation packaging system because transgene expression from the ltr is tat dependent. How can lentivirus be used to make stable cell lines.

Our solid expertise in lentiviral technology enables us to efficiently generate stable cell lines to meet the requirements of reporter cells that express rfp gfp luciferase for high throughput assays overexpression cells to study your protein of interest tet inducible cells or customized cells for any research application. Previous approaches to lentiviral vector producer cell line generation were typically based on sequential stable transfection or transduction of dna encoding each vector component transfer vector gagpol rev and vsvg into host cells at separate genomic loci. Lentiviruses can be used to make stable cell lines in the same manner as standard retroviruses. The technique of generating stable cell lines using 3rd generation lentivirus is very robust and it typically takes about 1 2 weeks to get stable expression for most mammalian cell lines.

They are used for gene down regulation by using shrna or for gene up regulation by using orf of the gene of interest. Unlike the short term protein expression observed using transient transfection approaches generating cell lines using lentiviral vectors enables long term protein expression studies. Shrna allows for stable knockdown of genes while sirna allows for transient knockdown. Lentiviral transduction to occur in my cells.

The cell line is created with stable transfection of bicistronic expression cassettes with re initiation of the translation mechanism.