For mammalian cells, of all the gene transfer methods available retroviral and lentiviral methods are great because they generate stably transduced cell lines with the constructive interest integrated into the cell genome.
But, particularly with some cells, getting your retroviral construct into your cell can be difficult, really difficult.
One way to approach this is to use Polybrene during your transduction step. But, Polybrene is cytotoxic. And after nurturing yourself so carefully, the last thing you want to do is kill of a whole bunch of them, right?
So, many researches end up finding themselves between a rock and a hard place.
Who really enjoys spending countless hours preparing lots of cells in retrovirus preps for transduction, throwing in cell killing additives like Polybrene, and hoping that you'll get enough candidate transductance? It's kind of like being in gene transfer purgatory.
But, with RetroNectin Reagent from Takara Bio, you can get high levels of gene transfer without using cytotoxic additives like Polybrene.
Here's how it works. Let's look in more detail at transduction using RetroNectin.
RetroNectin Reagent is a chimeric peptide of recombinant human fibronectin fragments. And it acts like a matchmaker between your retroviral particles and your cells. Retrovirus is hypothesized to associate with the heparin binding domain in the center on the RetroNectin molecule.
Transduction-inhibiting materials from the retroviral prep supernatant can be washed away, which further enhances gene transfer efficiency. Then target cells can be added.
Cells are mainly thought to bind through association of the VLA-4 receptor with the CS-1 domain and can also interact through the VLA-5 receptor and the RetroNectin RGDS domain.
By associating retroviral particles more efficiently with your cells, gene transfer rates improve dramatically, no need for Polybrene. And since the efficiency is higher, you can start with fewer cells.
The protocol is easy. You can coat plates yourself starting with lyophilized RetroNectin Reagent and even use pre-coated dishes that are ready to go.
There's simply no comparison for gene transfer rate. RetroNectin is better than Polybrene, better than Protamine.
RetroNectin has been used on dozens and dozens of cell types, including some that are really tough to work with, even primary cells.
But, don't take our word for it. RetroNectin Reagent has been used in over 1,700 peer-reviewed studies. And clinical grade RetroNectin Reagent has been used in over 57 gene therapy clinical trials at institutions all over the world.
Try Takara's RetroNectin Reagent today. To avoid gene transfer purgatory, treat your cells with tender love and care and move your research forward.
Find out more about RetroNectin Reagent at our Website. Visit www.clonetech.com/takara today.
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CNPG_Save-time-cells-by-enhancing-lentiviral-and-retroviral-transduction-efficiency