One cell model establishment to inhibit CaMKIIγmRNA expression in the dorsal root ganglion neuron by RNA interfere.
CaMKIIγin dorsal root ganglion neurons is closely related to the neuropathic pain, neuron injury induced by local anesthetics. To get great insight into the function of CaMKIIγin dorsal root ganglion neurons, we need one cell model to specially inhibit the CaMKIIγmRNA expression. The present study was aimed to establish one cell model to specially inhibit the CaMKIIγmRNA expression. We designed the CaMKIIγshRNA sequence and connected with pYr-1.1 plasmid. The ligation product of the CaMKIIγshRNA and pYr-1.1 plasmid was recombined with pAd/PL-DEST vector into pAD-CaMKIIγ-shRNA. adenovirus vector. pAD-CaMKIIγ-shRNA. adenovirus vector infected the dorsal root ganglion neuron to inhibit the CaMKIIγmRNA expression in vitro. The pAD-CaMKIIγ-shRNA adenovirus vector was verified to be correct by the digestion, sequence. And pAD-CaMKIIγ-shRNA. adenovirus vector can infect the DRG cells to inhibit the CaMKIIγmRNA or protein expression by the real-time polymerase chain reaction (PCR) or western blotting. Those results showed that we successfully constructed one adenovirus vector that can infect the dorsal root ganglion neuron to inhibit the CaMKIIγmRNA and protein expression. That will supply with one cell model for the CaMKIIγfunction study.
a Department of Anesthesiology , The First People's Hospital of Foshan and Foshan Hospital of Sun Yat-Sen University , Foshan , China.
Xianjie
X
Wen
a Department of Anesthesiology , The First People's Hospital of Foshan and Foshan Hospital of Sun Yat-Sen University , Foshan , China.|b Department of Pain Clinic , the First Affiliated Hospital of Jinan University , Guangzhou , China.
Xiaohong
X
Li
a Department of Anesthesiology , The First People's Hospital of Foshan and Foshan Hospital of Sun Yat-Sen University , Foshan , China.
Hua
H
Liang
a Department of Anesthesiology , The First People's Hospital of Foshan and Foshan Hospital of Sun Yat-Sen University , Foshan , China.
Chenxiang
C
Yang
a Department of Anesthesiology , The First People's Hospital of Foshan and Foshan Hospital of Sun Yat-Sen University , Foshan , China.
Jiying
J
Zhong
a Department of Anesthesiology , The First People's Hospital of Foshan and Foshan Hospital of Sun Yat-Sen University , Foshan , China.
Hanbing
H
Wang
a Department of Anesthesiology , The First People's Hospital of Foshan and Foshan Hospital of Sun Yat-Sen University , Foshan , China.
Hongzhen
H
Liu
D000256:Adenoviridae; D000818:Animals; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000096:biosynthesis* / Q000235:genetics; D005727:Ganglia, Spinal / Q000166:cytology / Q000201:enzymology*; D015971:Gene Expression Regulation, Enzymologic; D008954:Models, Biological; D009474:Neurons / Q000166:cytology / Q000201:enzymology*; D034622:RNA Interference; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics; D034741:RNA, Small Interfering / Q000096:biosynthesis* / Q000235:genetics; D051381:Rats; D014161:Transduction, Genetic
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; C517678:Camk2g protein, rat
Dorsal root ganglion neurons; RNA interfere; calcium/calmodulin-dependent protein kinase II gamma; cell culture; neuroscience