Development and evaluation of a novel nano-scale vector for siRNA.
To synthesize a lipid-cationic polymer (LCP) containing brassidic acid side chain and to investigate its transfection efficiency and characteristics as a siRNA gene vector. The LCP was chemically synthesized and its nucleic acid binding capacity was determined by gel electrophoresis. HeLa-EGFP and TH1080-EGFP cell lines were transfected with siRNA against enhanced green fluorescent protein (EGFP) gene using a LCP to investigate the transfection efficiency. An MTT assay was performed to evaluate the cellular toxicity of the LCP vector. Its degradability and stability under acidic conditions were also investigated. The LCP vector possessed high DNA binding capacity. More than 73% of the cellular fluorescence was inhibited by the LCP-mediated transfection of siRNA against EGFP gene, indicating that vector had high transfection efficiency. Cellular viability was about 95% at the optimum transfection efficiency of LCP, suggesting that the cellular toxicity of LCP was very low. The LCP was also observed to be degradable; moreover, it could be easily stored at normal temperature. A gene vector used for the transfection of siRNA was successfully fabricated from synthesized LCP. Its numerous excellent properties entitle values for further scientific research.
Jing
Guang-Jun
GJ
Center of Clinical Laboratory, Zhongshan Hospital, Medical College of Xiamen University, Xiamen 361004, China.
Fu
Zuo-Gen
ZG
Dan
Bing
B
Lin
Li-Rong
LR
Yang
Tian-Ci
TC
Shi
Song-Lin
SL
D002021:Buffers; D002412:Cations; D016923:Cell Death; D002470:Cell Survival; D004247:DNA / Q000378:metabolism; D003851:Deoxyribonucleases / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D005453:Fluorescence; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D008055:Lipids / Q000737:chemistry; D049329:Nanostructures / Q000737:chemistry*; D009711:Nucleotides / Q000378:metabolism; D010316:Particle Size; D011108:Polymers / Q000737:chemistry; D034741:RNA, Small Interfering / Q000378:metabolism*; D044967:Serum; D055672:Static Electricity; D013696:Temperature; D017186:Titrimetry; D014162:Transfection / Q000379:methods*
D002021:Buffers; D002412:Cations; D008055:Lipids; D009711:Nucleotides; D011108:Polymers; D034741:RNA, Small Interfering; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D004247:DNA; D003851:Deoxyribonucleases