Stable Cell Line Generation Lentivirus

They are used for gene down regulation by using shrna or for gene up regulation by using orf of the gene of interest. The authors describe a new lentiviral vector packaging cell line lentipro26 using a mutated less active viral protease to overcome potential cytotoxic limitations. The cell line is created with stable transfection of bicistronic expression cassettes with re initiation of the translation mechanism.

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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.

Lentiviruses can be used to make stable cell lines in the same manner as standard retroviruses. 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. 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. Shrna allows for stable knockdown of genes while sirna allows for transient knockdown.

How can lentivirus be used to make stable cell lines. The advantage of using the 3rd generation lentivirus are that are very safe and they are replication incompetent. Lentiviruses are used widely to generate stable expression mammalian cell lines. Thus we believe that this study demonstrates that transduction of packaging lines by csin.

That is many lentiviral genomes. 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. All 2nd generation lentiviral transfer plasmids must be used with a 2nd generation packaging system because transgene expression from the ltr is tat dependent. Lentiviral vectors offer stable integration of your gene of inte.

Unlike the short term protein expression observed using transient transfection approaches generating cell lines using lentiviral vectors enables long term protein expression studies. Furthermore a third generation lentivirus vector stable packaging cell line deficient for the hiv 1 tat gene was successfully used to generate high titer csin vector stocks following transduction with the ptk136 csin vector kafri unpublished data. 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. Lentiviral transduction to occur in my cells.

Generation of stable cell lines using lentivirus.

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