One cell model establishment to inhibit CaMKII_γ mRNA expression in the dorsal root ganglion neuron by RNA interfere

Abstract

CaMKII_γ in 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 CaMKII_γ in dorsal root ganglion neurons, we need one cell model to specially inhibit the CaMKII_γ mRNA expression. The present study was aimed to establish one cell model to specially inhibit the CaMKII_γ mRNA expression. We designed the CaMKII_γ shRNA sequence and connected with pYr-1.1 plasmid. The ligation product of the CaMKII_γshRNA 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 CaMKII_γ mRNA 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 CaMKII_γ mRNA 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 CaMKII_γ mRNA and protein expression. That will supply with one cell model for the CaMKII_γ function study.

Keywords:

• Dorsal root ganglion neurons
• calcium/calmodulin-dependent protein kinase II gamma
• RNA interfere
• neuroscience
• cell culture

Disclosure statement

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of this article.