Construction of lentiviral recombinants expressing shRNA targeting N-methyl-D-aspartate gene

Objective N-methyl-D-aspartate in anterior cingulate cortex is relevant to the dispirital mood of cancer patients with pain.The aim of this study is to construct shRNAs-expressing lentiviral recombinants targeting the rat N-methyl-D-aspartate(NR2B) gene.Methods Firstly the shRNAs-expressing lentiviral recombinants targeting the rat NR2B gene were constructed and then identified by PCR analysis.The shRNAs-expressing lentiviral recombinants targeting the rat NR2B gene were then transfected into 293T cells with the NR2B gene-expressing plasmid.Western blot was employed to assess the gene silencing efficacy of these recombinants.Results PCR analysis and sequencing showed that shRNAs were correctly inserted into the lentiviral vector and three recombinants were constructed successfully.All of these three recombinants could achieve gene knockdown effects,and the recombinants Ⅲ had the most significant gene silencing effect among them.Conclusion shRNAs-expressing lentiviral recombinants targeting the NR2B gene have been successfully constructed and can be used in transgenic analgesic research.