Research Of SiRNA Mediated By Non-virus Vector

RNAi is mediated by the non-virus vector,dsRNA is mediated into the cancer cells,result in breaking down the mRNA,which make relevant gene not express.so the process is also called gene silencing.It is one of the most briliant methods in gene therapy of genatic levels.Currently,this method is generally used in micology.zoology and botany.The advantage of RNAi is that has better characteristic,use simplely and conviniently,transfer efficiently so and on.Now that it is widely used as estimating the function of gene,gene therapy and medical selection,which exert important effect.Through the use of small interfering RNA whose size is seperated to21-25nucleotides from RNAi perevents some special gene expressing.This dessertation renewed the gene threapy,gene vector,liposome,the inner process of the component of cationic liposome and DNA,RNAi,siRNA,plasmid.And select the cationic liposome C/D and Lipofectamine2000as gene vector;through using the luciferase gene do the experiment of RNAi.Next follows the main process of experiment and consequence.C/D and Lipofectamine2000transferred the plasmid DNA into the Hela cells,observing the efficiency of transfection. Secondly.researched more about the delaying reaction of the component between cationic liposome and siRNA;in addition,Different concentrations siRNA were used to silence luciferase gene’s activity,the result was examined by microplate reader.And MTT assay was used to analyze the cells viability which were transfected.As a result, C/D and Lipofectamine2000could efficiently transfer the plasmid DNA,and had a binding ability with siRNA.The lower concentration of siRNA.expressed the high efficent in silencing level,and affected little to cell viability.The survival rate of cells were decreasing while the concerntration of siRNA was inceasing.In conclusion, cationic liposome C/D and Lipofectamine2000can transfer a little lower concentration siRNA to silence target gene’s activity efficiently.Cationic liposome C/D can silence luciferase gene more efficiently,but cause a higer cell Cytotoxicity.By optimizing the experimental conditions,cationic liposome can transfer low concentration siRNA to silence target gene’s activity efficiently.