Antisense inhibition of delta-opioid receptor gene function in vivo by peptide nucleic acids.
Antisense inhibition of delta-opioid receptor gene function in vivo by peptide nucleic acids.
复制标题
肽核酸对体内δ-阿片受体基因功能的反义抑制。
作者：
G. Fraser;J. Holmgren;P. Clarke;C. Wahlestedt
肽核酸（PNA）是DNA的合成类似物，其与互补的寡核苷酸序列杂交，具有优异的亲和力和靶特异性。PNA在生物液体中的稳定性以及这些结构的独特杂交特性表明PNA可能具有相当大的潜力作为体内实验使用的反义试剂。为了验证这一假设，我们试图使用PNA序列来调节大鼠脊髓上δ阿片受体的功能，PNA序列被设计为与大鼠δ阿片受体的一个区域互补。重复i. c. v.管理PNA超过5天的时间显着抑制的抗伤害性反应和运动反应的选择性δ阿片受体激动剂。PNA以序列特异性、靶特异性和可逆的方式减弱δ-阿片受体功能，这是由反义机制引起的功能抑制的特征。基于动物的行为和对处理组织的检查，PNA处理没有引起明显的毒性。脑匀浆的饱和结合研究未显示给药组之间受体B（max）存在任何显著差异。然而，[（35）S]鸟苷-5 '-O-（3-硫代）三磷酸结合试验表明，在从反义处理大鼠制备的匀浆中，激动剂效力显著降低。总之，这些结果证明肽核酸是体内有效的反义试剂，并表明PNA可以是磷酸二酯或硫代磷酸酯寡核苷酸或其变体的有用替代物，用于体内基因功能的测定。
Peptide nucleic acids (PNA) are synthetic analogs of DNA that hybridize to complementary oligonucleotide sequences with exceptional affinity and target specificity. The stability of PNA in biological fluids together with the unique hybridization characteristics of these structures suggests that PNA may have considerable potential as antisense agents for experimental use in vivo. To test this hypothesis, we attempted to modulate supraspinal delta-opioid receptor function in rats using PNA sequences designed to be complementary to a region of the rat delta-opioid receptor. Repeated i.c.v. administration of PNA over a period of 5 days significantly inhibited the antinociceptive response and locomotor response to selective delta-opioid receptor agonists. PNA attenuated delta-opioid receptor function in a sequence-specific, target-specific, and reversible manner characteristic of the functional inhibition caused by an antisense mechanism. There were no apparent toxicities arising from the PNA treatment based on the behavior of the animals and inspection of the treated tissues. Saturation binding studies on brain homogenates did not reveal any significant difference in receptor B(max) between treatment groups. However, [(35)S]guanosine-5'-O-(3-thio)triphosphate binding assays demonstrated a significant decrease in agonist efficacy in homogenates prepared from antisense-treated rats. Taken together, these results demonstrate that peptide nucleic acids are effective antisense agents in vivo and suggest that PNA may be a useful alternative to phosphodiester or phosphorothioate oligonucleotides, or variants thereof, for determination of gene function in vivo.
反义寡脱氧核苷酸介导的体内阿片受体“敲低”后阿片受体选择性激动剂抗伤害作用的表征。
DOI：
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发表时间：
1996
期刊：
The Journal of pharmacology and experimental therapeutics.
影响因子：
--
作者：
Bilsky,EJ;Bernstein,RN;Hruby,VJ;Rothman,RB;Lai,J;Porreca,F
通讯作者：
Porreca,F
D2 多巴胺受体反义寡脱氧核苷酸抑制 D2 多巴胺受体功能池的合成。
DOI：
--
发表时间：
1995
期刊：
Molecular pharmacology.
影响因子：
--
作者：
Qin,ZH;Zhou,LW;Zhang,SP;Wang,Y;Weiss,B
通讯作者：
Weiss,B
影响因子：
56.9
作者：
EVANS, CJ;KEITH, DE;EDWARDS, RH
通讯作者：
EDWARDS, RH