By SIRION Biotech…
SIRION enhancers for viral transduction
SIRION Biotech has developed its unique BOOST™ products to specifically enhance lentiviral and adenoviral transduction efficiency during gene delivery into difficult to transduce mammalian and rodent cells, significantly increasing virus uptake without toxicity.
Transduction with lentivirus and adenovirus
Transduction of virus particles to deliver genes to a cell is inherently a highly efficient wa to genetically modify mammalian or human tissue. However, in some circumstances transduction needs to be optimized to overcome specific limitations.
Lentiviral vector transduction works to introduce foreign DNA into another cell to generate stable transgene expression or gene knockdown, whereas adenoviral transduction leads to transient gene expression or knockdown. Measuring protein expression, typically by Western blot or immunostaining, can validate presence of the newly introduced genetic material.
Vector optimization strategies
Apart from identifying the most appropriate vector system, the SIRION lentivirus development service offers various ways to achieve higher transduction or viral transfection levels in gene transfer.
The first is through use of transduction enhancers – reagents added to the transduction medium to increase success rates.
The cationic polymer polybrene can be used independent of receptor biologies to improve the general physicochemical interactions taking place between cell membrane and virion during transduction. Small polybrene/lentivirus concentrations have been shown to yield 10-fold increases in transduction as well as significant increase in virus adsorption rates on murine fibroblasts, independent of receptor and envelope properties.
SIRION offers proprietary transduction enhancers for preclinical and clinical application of lentiviral vectors and preclinical application of adenoviral vectors. These transduction boosters are reagents that are added to the transduction medium to increase success rates, working as adapters between the cell- and virus receptors to enable better adherence to cells that carry a fitting receptor-makeup and increase the uptake of the virus.
- LentiBOOST® is a highly effective, non-cytotoxic and universally acting (receptor independent) adjuvant that can be applied to a wide range of clinically relevant cell types including CD34+ hematopoietic stem cells (HSCs), primary T cells and NK cells. These unique features make it a promising candidate to improve clinical transduction protocols for ex vivo gene therapies and CAR-T cell therapies. Benefits include up to 90% improvement in lentiviral transduction efficiency, Increased expression levels of therapeutic protein with favorable effect on clinical success rates, positive impact on cell proliferation for CD34+ and T cells, proven lack of cell toxicity and reduced cost of transgenic cell manufacturing.
- AdenoBOOST™ is a non-toxic peptide-based solution added to adenovirus particles prior to transduction to specifically bridge the adenovirus surface to the cell membrane, thereby enhancing gene expression by 20 to 50-fold into cell types lacking the primary adenovirus CAR-receptor.
Different cell types exhibit varying susceptibilities to virus particles. Often, small changes to an existing lentiviral transduction protocol, such as increasing viral load or incubation time can have a significant effect. Added steps can further help maximize the impact. A study has found that integrating a centrifugation step into lentiviral transduction can almost double the proportion of transduction-positive SUDHL-1 cells to more than 80% (Anastasov et al, 2016).
As the technology expert and service provider for vector customization, SIRION Biotech has collected and optimized transduction protocols for more than 150 pairings between cell types and viruses.
Click on SIRION Biotech Transduction enhancers for more information.