[Construction of siRNA expression vector of NgR and its inhibition on the expression of NgR].
OBJECTIVE
To construct the expressing vector of siRNA in order to inhibit NgR expression, which provides a new gene therapy approach to the optic nerve impairment.


METHODS
It was a experimental study. Specific short chain oligonucleotides was designed by using the siRNA software according to the mRNA sequence provided by genebank, the double chain DNA sequence was gained through annealing after chemosynthesis and was inserted to the pSUPER-EGFP vector linearized with Bgl II and Hind II enzyme, the recombinant expression vector was evaluated by using EcoR I and Hind II enzyme cutting and sequencing. At last, the constructed vectors were transfected into wistar rat retinal ganglion cells and the expression level of NgR was observed.


RESULTS
Identification by enzyme cutting and sequencing showed that the expression vector was constructed successfully, and it was proved that the NgR expression level was effectively inhibited.


CONCLUSIONS
The constructed siRNA expression vectors can block the NgR gene expression, it will be the foundation for the research of gene therapy to optic nerve regeneration.
Chen
Chun-Lin
CL
Research Institute of Field Surgery, Da Ping Hospital, Third Military Medical University, Chongqing 400042, China.
Chen
Xiao-Fan
XF
Yin
Xiao-Lei
XL
Ye
Jian
J
D000818:Animals; D015870:Gene Expression; D005822:Genetic Vectors; D009185:Myelin Proteins / Q000235:genetics*; D034741:RNA, Small Interfering; D051381:Rats; D017208:Rats, Wistar; D011956:Receptors, Cell Surface / Q000235:genetics*; D014162:Transfection
D009185:Myelin Proteins; D034741:RNA, Small Interfering; D011956:Receptors, Cell Surface