Glycofect™ Transfection Reagent

Glycofect is a new DNA carrier that elicits low cytotoxic response coupled with high delivery efficiency and transgene expression.  Glycofect is a superior reagent for primary cell transfection as compared to leading lipid and poly-cationic based products. 

Features

  

• Effective delivery with low cytotoxicity

• Effective cellular uptake through nanoparticle delivery

• Protects DNA from nuclease degradation

• Biodegrades after endocytosis

• Simple & robust transfection procedure

• Exceptional results in primary cells

Biodegradable Release Process

Glycofect is a cationic carbohydrate-containing synthetic polymer based on the poly-glycoamidoamine (PGAA) polymer library developed in the group of world-renowned biomaterials professor Theresa M. Reineke.  When formulated with a nucleic acid, the polymer compacts the genetic material into a positively charged nanoparticle (dh = ~100 nm) suitable for cellular uptake.  The distinguishing trait of Glycofect which gives it low cytotoxicity and high delivery efficiency is that it biodegrades in cytoplasm-like conditions thereby promoting gene release after the DNA-Glycofect particle has been endocytosed.  This timed release process allows the genetic material being carried by the particle to be fully protected and shielded from degradation and/or immune response until it is released in the proximity of the cell nucleus where it may be expressed.  Indeed, Glycofect promotes low toxicity transfections in immortalized cells, constantly outperforming lipid-based competitors.  However the arena where Glycofect truly outperforms is in transfecting primary cells, where low cytotoxic response is especially important due to the expense and labor required to obtain these fragile cell types.Because Glycofect is biodegradable in nature, it is packaged in separate single-use aliquots, each suitable for transfection of one 24-well plate (easily adapted to other plating formats).

Documented Results

 

The following micrographs compare Glycofect to leading lipid-based reagents when used to transfect primary dermal fibroblasts and primary aortic smooth muscle cells. In each series, the DIC images (top row) show the density of the individual cells while the fluorescence iimages (bottom row) show those cells that have been transfected with a plasmid expressing green fluorescent protein. All samples were prepared per manufacturers' instructions.  

                                      Primary Dermal Fibroblasts

                  Control                                   Glycofect                             Lipid Competitor A                   Lipid Competitor B

                                Primary Human Aortic Smooth Muscle

                  Control                                   Glycofect                             Lipid Competitor A                   Lipid Competitor B

Technical References

  • T.M. Reineke "PGAAs: " Cationic Glycopolymers for DNA Delivery" J. Polym. Sci. A: Polym. Chem. 2006, 44, 6895-6908.
  • Y. Liu, T.M. Reineke, "Poly(glycoamidoamine)s for Gene Delivery.  Stability of Polyplexes and Efficacy with Cardiomyoblast Cells" Bioconjugate Chem. 2006, 17, 101-108.
  • Y. Liu, T.M. Reineke, "Degradation of Poly(glycoamidoamine) DNA Delivery Vehicles:  Polyamide Hydrolysis at Physiological Conditions Promotes DNA Release."  Biomacromolecules, 2010, 11, 316.
  • Y. Liu, T.M. Reineke, "Hydroxyl Stereochemistry and Amine Number wihin Poly(glycoamidoamine)s Affect Intracellular DNA Delivery."  J. Am. Chem. Soc., 2005, 127, 3004

Resource Links

Glycofect Fact Sheet              Glycofect Protocol              Glycofect MSDS