SAFETRANS (ƒ¿-CDE)


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Background

Gene and oligonucleotide therapies are emerging as a potential strategy for the treatment of genetic diseases, cancers, cardiovascular diseases and infectious diseases. Recently, polyamidoamine (PAMAM) dendrimer (generation (G) 3) conjugate with α-cyclodextrin (α-CDE) has been known as novel DNA, shRNA and siRNA carriers. α-CDE works as a superior nucleic acids carrier without severe cytotoxicity in vitro and in vivo.

Protocol

1. Seed the cells (1 x 104 ~2 x 105 cells/well) on a 24-well-plate, and cultured for 1 day at 5% CO2 incubator. (40~60 % confluent should be set for transfection.)
2. Add the serum-free medium 200 uL/well into 1.5 mL tube, and add the pDNA (2 ug/well) dissolved in TE buffer (pH 7-8)
3. Add the α-CDE Transfection reagent (1 mg/mL) 189.3 uL into pDNA solution of 2 and mixing by pipetting (5~6 times) or slight vortexing (10 s).
4. Incubation for 10-15 min at room temperature to form the complex with pDNA.
5. During the complexation in 4, cells were washed with PBS or serum free medium.
6. Add the serum-free medium 200 uL/well and pDNA complex solution (1 ug/uL) prepared in 3, then incubated for 1-3 h at 5% CO2 incubator 1) .
7. After removal of the medium containing pDNA complex, washed with PBS or serum-free medium for 1 time.
8. Add the culture medium (containing serum) and further incubated for 24 h, and determine the gene expression.

References

1. Kihara F, Arima H, Tsutsumi T, Hirayama F, Uekama K. In Vitro and In Vivo Gene Transfer by an Optimized ƒ¿-Cyclodextrin Conjugate with Polyamidoamine Dendrimer. Bioconjug. Chem., 14, 342-350, 2003.
2. Kihara F, Arima H, Tsutsumi T, Hirayama F, Uekama K. Effects of Structure of Polyamidoamine Dendrimer on Gene Transfer Efficiency of the Dendrimer Conjugate with ƒ¿-Cyclodextrin. Bioconjug. Chem., 13, 211-1219, 2002.
3. Arima H, Kihara F, Hirayama F, Uekama K. Enhancement of Gene Expression by Polyamidoamine Dendrimer Conjugates with ƒ¿-, ƒÀ-, ƒÁ-Cyclodextrins. Bioconjug. Chem., 12, 476-484, 2001.
4. Tsutsumi T, Hirayama F, Uekama K, Arima H. Potential use of Polyamidoamine Dendrimer/ƒ¿-Cyclodextrin Conjugate (generation 3, G3) as a Novel Carrier for Short Hairpin RNA-Expressing Plasmid DNA. J. Pharm. Sci., 97, 3022-3034, 2008.
5. Tsutsumi T, Hirayama F, Uekama K, Arima H. Evaluation of polyamidoamine dendrimer/ƒ¿-cyclodextrin conjugate (generation 3, G3) as a novel carrier for small interfering RNA (siRNA). J. Control. Release, 119, 349-359, 2007.
6. Tsutsumi T, Hirayama F, Uekama K, Arima H. Potential use of polyamidoamine dendrimer/ƒ¿-cyclodextrin conjugate (generation 3, G3) as a novel carrier for short hairpin RNA-expressing plasmid DNA, J. Pharm. Sci., 97, 3022-3034, 2008.
7. Anno T, Motoyama K, Higashi T, Hirayama F, Uekama K, Arima H. Preparation and evaluation of polyamidoamine dendrimer (G2)/branched-ƒÀ-cyclodextrin conjugate as a novel gene transfer
carrier. J. Incl. Phenom. Macrocycl. Chem., in press, 2010.

Product List

Product Name Cat# Quantity Price

SAFETRANS ((α)-CDE) DataSheet ,

CSR-KMU-T01

2 MG N/A

SAFETRANS (GUG-(β)-CDE) DataSheet ,

CSR-KMU-T02

2 MG N/A

SAFETRANS (Lac-(α)-CDE) DataSheet ,

CSR-KMU-T03

2 MG N/A

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.