Cationic liposome for transfection reagents, preparation and application thereof
A technology of cationic liposomes and transfection reagents, which is applied in liposome delivery, medical preparations of non-effective ingredients, and the use of microcapsules. It can solve the problems of high price and low transfection efficiency, and achieve low price, The effect of high transfection efficiency
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0026] Transfection reagent prepared according to the present invention, its molecular formula is (A):
[0027]
[0028] The synthetic route is:
[0029]
[0030] 1) Synthesis of compound I
[0031] In an Erlenmeyer flask, 40 mL of glacial acetic acid was added to benzotriazole (3.8 g; 31.8 mmol). Slowly add 50 mL of bleach to the solution. The mixture was stirred at room temperature for 2 h. The resulting white solid was filtered and washed with water until pH=7 of the filtrate. Drying under vacuum afforded 1-chlorobenzotriazole as a white solid in 90% yield (4.38 g). Under nitrogen protection, benzotriazole (2.4 g; 20 mmol) and 1-chlorobenzotriazole (3.07 g; 20 mmol) were dissolved in 100 mL of dichloromethane. The mixture was cooled to -80°C, and 15 mL of dichloromethane containing n-dodecanethiol (5.06 g; 25 mmol) was slowly added dropwise into the solution. After stirring at -80°C for 1 h, a suspension in THF containing 10 mL of thiourea (1.53 g; 20 mmol) was ...
Embodiment 2
[0041] Use commercially available cationic lipid transfection reagent LipofectAMINEPLUS, FuGENE TM -6 is compared with compounds of the present invention. HEK-293 (human embryonic kidney-derived cells), COS-7 (African green monkey kidney fibroblasts transformed with SV40 virus gene) and HeLa (human cervical cancer-derived cells) with β-galactosidase reporter plasmid DNApCMV·SPORT-β -gal (Life Technologies Ltd.) for the reaction.
[0042] 1) One day before transfection, cells were seeded in a 24-well tissue culture plate containing 0.4 ml of culture medium. The medium is DMEM medium containing 1% non-essential amino acids and 10% fetal bovine serum. For HEK-293 and COS-7 cell lines, coat the plate with Poly-L-Lysine before inoculation to increase cell adhesion.
[0043] 2) On the second day, the DNA transfection reagent was prepared as follows:
[0044] The cationic lipid reagent and DNA were diluted to 25ul respectively with serum-free DMEM containing 1% NEAA. For Lipofec...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More