Silencing GIRK4 expression in human atrial myocytes by adenovirus-delivered small hairpin RNA.
GIRK4 has been shown to be a subunit of I(KACh), and the use of GIRK4 in human atrial myocytes to treat arrhythmia remains an important research pursuit. Adenovirus-delivered small hairpin RNA (shRNA) has been used to mediate gene knockdown in mouse cardiocytes, yet there is no information on the successful application of this technique in human cardiocytes. In the current study, we used a siRNA validation system to select the most efficient sequence for silencing GIRK4. To this end, adenovirus-delivered shRNA, which expresses this sequence, was used to silence GIRK4 expression in human atrial myocytes. Finally, the feasibility, challenges, and results of silencing GIRK4 expression were evaluated by RT-PCR, western blotting, and the voltage-clamp technique. The levels of mRNA and protein were depressed significantly in cells infected by adenovirus-delivered shRNA against GIRK4, approximately 86.3% and 51.1% lower than those cells infected by adenovirus-delivered nonsense shRNA, respectively. At the same time, I(KACh) densities were decreased 53% by adenovirus-delivered shRNA against GIRK4. In summary, adenovirus-delivered shRNA against GIRK4 mediated efficient GIRK4 knockdown in human atrial myocytes and decreased I(KACh) densities. As such, these data indicated that adenovirus-delivered shRNA against GIRK4 is a potential tool for treating arrhythmia.
Liu
Xiongtao
X
Department of Cardiology, Affiliated Tangdu Hospital of the Fourth Military Medical University, Xi'an, 710038, People's Republic of China.
Yang
Jian
J
Shang
Fujun
F
Hong
Changming
C
Guo
Wangang
W
Wang
Bing
B
Zheng
Qiangsun
Q
D000256:Adenoviridae / Q000235:genetics; D001145:Arrhythmias, Cardiac / Q000628:therapy; D051676:G Protein-Coupled Inwardly-Rectifying Potassium Channels / Q000032:analysis / Q000235:genetics*; D020868:Gene Silencing; D006325:Heart Atria / Q000166:cytology; D006801:Humans; D032383:Myocytes, Cardiac / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000494:pharmacology
D051676:G Protein-Coupled Inwardly-Rectifying Potassium Channels; C484274:KCNJ5 protein, human; D034741:RNA, Small Interfering