Construction of RNAi vector of dopamine D1 receptor and identification of its silencing effects.
10.3969/j.issn.1672-7347.2013.06.004
- Author:
Hui LI
1
;
Junmei XU
;
Guixiu YUAN
;
Jin LI
;
Zhangling CHEN
Author Information
1. Department of Anesthesiology, Central South University, Changsha, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Cell Line, Tumor;
Genetic Vectors;
Glioma;
pathology;
Hybrid Cells;
Liposomes;
metabolism;
Mice;
Neuroblastoma;
pathology;
RNA Interference;
RNA, Messenger;
genetics;
metabolism;
RNA, Small Interfering;
genetics;
Receptors, Dopamine D1;
genetics;
metabolism;
Transfection
- From:
Journal of Central South University(Medical Sciences)
2013;38(6):570-575
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To construct dopamine D1 receptor (DRD1) expression interference vectors to study the role of DRD1 in nerve cells and lay a foundation for drug development in anti-convulsion.
METHODS:Based on DRD1 gene sequence in GenBank, 10 interfere vectors of DRD1 were designed. Liposomal was used to transfect NG-108-15 and the transfect effect was assayed by GFP. With realtime PCR and Western blot, the DRD1 expression was detected.
RESULTS:The 10 constructed interfere vectors transfected into NG-108-15 cells by liposomal method and inhibited DRD1 mRNA and protein expression. DRD1 mRNA expression in NG-108-15 cells transfected with pGPU6-GFP-Neo-si-DRD1-5 was the lowest whereas DRD1 protein expression in NG-108-15 cells transfected with pGPU6-GFP-Neo-si-DRD1-1, -2, -6, -7 was the lowest.
CONCLUSION:DRD1 expression interference vector is successfully constructed.
