Abstract: Objective To construct Neuralized siRNA vector and to observe the effect of siRNA plasmids on expression of Neuralized in cells. Method Short hairpin oligonucleotide sequence producted by chemical synthesis was first inserted into linearized plasmid. After indentificated by enzymatic digestion and sequencing, Neuralized siRNA plasmids were transfected into HEK293 cells, the expression level and change of location of Neuralized were analysis by Western-Blot and Immunofluorescence. Result Sequencing result from Neuralized siRNA vector showed that the target nucleotide sequences were successfully inserted into the expected sites of vectors and sequences were correct. After the plasmids were transfected HEK293 cells, Western-Blot displayed that neuralized expression was decreased in cells. Immunofluorescence displayed that the fluorescence of neuralized lacted on the cell membrane was dimmed when the cells were transfected the four kinds of neuralized siRNA plasmids. Conclusion The neuralized siRNA plasmids were successfully constructed and could decrease expression level of neuralized in cells.

Key words: Neuralized, SiRNA-interference