TY - JOUR
T1 - Targeting neuronal nitric oxide synthase by a cell penetrating peptide Tat-LK15/siRNA bioconjugate
AU - Peng, Jie
AU - Rao, Yun
AU - Yang, Xue
AU - Jia, Ji
AU - Wu, Youping
AU - Lu, Jianhua
AU - Tao, Yuanxiang
AU - Tu, Weifeng
N1 - Funding Information:
This work was funded by the National Natural Science Foundation of China (81100817 and 81371233) and Science and Technology Project of Guangdong Province (2014A020212555). The authors would like to thank laboratory of Guangzhou General Hospital of Guangzhou Military Command for their technical support and Professor Yuan-Xiang Tao of New Jersey Medical School for helpful discussion on projects.
Publisher Copyright:
© 2017 Elsevier B.V.
PY - 2017/5/22
Y1 - 2017/5/22
N2 - We developed a cell penetrating peptide (CPP) Tat-LK15, as a siRNA carrier to target nNOS. The feasibility, stability, efficiency and selectivity of this peptide-siRNA complex were evaluated in rat neuronal cells. We also compared the new method with conventional siRNA carrier Lipofectamine™. It was found that the CPP Tat-LK15 effectively and specifically delivered nNOS-siRNA into Rat retinal ganglia (RGC-5) cells and silenced the expression of nNOS. The CPP Tat-LK15 can conjugate with siRNA to form stable complex at a ratio of 2:1 (peptide/siRNA, w/w), which maintained stable in serum for as long as 4 h. The CPP Tat-LK15 was low-toxicity to cells, as the apoptosis rate of treat cells was not increased significantly when the used peptide lower than 10 μg/mL. Moreover, the cellular uptake of nNOS siRNA by Rat Neurons-dorsal spinal cord (RNdsc) cells was also significantly more than naked siRNA by RNdsc cells. The CPP Tat-LK15 was an efficient and stable, and non-cytotoxic siRNA delivery to neurons and effectively silenced the nNOS expression. The CPP Tat-LK15 mediated siRNA delivery was a potential tool to treat neuropathic diseases involving NO or nNOS neurotoxic cascades.
AB - We developed a cell penetrating peptide (CPP) Tat-LK15, as a siRNA carrier to target nNOS. The feasibility, stability, efficiency and selectivity of this peptide-siRNA complex were evaluated in rat neuronal cells. We also compared the new method with conventional siRNA carrier Lipofectamine™. It was found that the CPP Tat-LK15 effectively and specifically delivered nNOS-siRNA into Rat retinal ganglia (RGC-5) cells and silenced the expression of nNOS. The CPP Tat-LK15 can conjugate with siRNA to form stable complex at a ratio of 2:1 (peptide/siRNA, w/w), which maintained stable in serum for as long as 4 h. The CPP Tat-LK15 was low-toxicity to cells, as the apoptosis rate of treat cells was not increased significantly when the used peptide lower than 10 μg/mL. Moreover, the cellular uptake of nNOS siRNA by Rat Neurons-dorsal spinal cord (RNdsc) cells was also significantly more than naked siRNA by RNdsc cells. The CPP Tat-LK15 was an efficient and stable, and non-cytotoxic siRNA delivery to neurons and effectively silenced the nNOS expression. The CPP Tat-LK15 mediated siRNA delivery was a potential tool to treat neuropathic diseases involving NO or nNOS neurotoxic cascades.
KW - Cell penetrating peptides
KW - Neuronal nitric oxide synthase (nNOS)
KW - Neuropathic pain
KW - RNA interference
KW - Small interfering RNA
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U2 - 10.1016/j.neulet.2017.04.045
DO - 10.1016/j.neulet.2017.04.045
M3 - Article
C2 - 28450191
AN - SCOPUS:85018296726
VL - 650
SP - 153
EP - 160
JO - Neuroscience Letters
JF - Neuroscience Letters
SN - 0304-3940
ER -