Repression of nicotinic acetylcholine receptor expression by antisense RNAs and an oligonucleotide.
Repression of nicotinic acetylcholine receptor expression by antisense RNAs and an oligonucleotide.
复制标题
通过反义 RNA 和寡核苷酸抑制烟碱乙酰胆碱受体的表达。
DOI：
10.1073/pnas.85.4.1302
复制
发表时间：
1988
影响因子：
11.1
通讯作者：
Miledi,R
中科院分区：
文献类型：
--
作者：
Sumikawa,K;Miledi,R
从电鳐电蚀斑乙酰胆碱受体4个亚基的cDNA克隆中合成4种反义RNA，研究它们对爪蟾卵母细胞中电鳐乙酰胆碱受体表达的影响。所有反义RNA抑制功能受体的出现在卵母细胞的表面膜至少1周。这种抑制是特异性的，因为反义RNA不阻断Cl-通道的表达，也由电鳐电斑mRNA编码。不完全反义RNA和合成寡核苷酸的实验表明，覆盖核糖体结合位点或mRNA中的起始密码子不是有效阻断的必要条件。因此，使用反义RNA结合爪蟾卵母细胞系统提供了一种新的方法来筛选cDNA文库的基因编码的多亚基神经递质受体。
Four antisense RNAs, synthesized from cDNA clones coding for the four subunits of the acetylcholine receptor of Torpedo electroplaques, were used to study their effect on the expression of functional Torpedo acetylcholine receptors in Xenopus oocytes. All antisense RNAs inhibited the appearance of functional receptors in the oocyte's surface membrane for at least 1 week. This inhibition was specific because the antisense RNAs did not block the expression of the Cl- channels, also encoded by Torpedo electroplaque mRNA. Experiments with incomplete antisense RNAs and a synthetic oligonucleotide indicate that covering the ribosome binding site or the initiation codon in the mRNA is not a necessary requirement for efficient blocking. Thus, the use of antisense RNAs combined with the Xenopus oocyte system provides a novel approach to screen cDNA libraries for the genes coding for multisubunit neurotransmitter receptors.