Antisense RNA complementary to 3' coding and noncoding sequences of creatine kinase is a potent inhibitor of translation in vivo.
Antisense RNA complementary to 3' coding and noncoding sequences of creatine kinase is a potent inhibitor of translation in vivo.
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与肌酸激酶 3 编码和非编码序列互补的反义 RNA 是体内翻译的有效抑制剂。
DOI：
10.1073/pnas.86.24.10006
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发表时间：
1989
影响因子：
11.1
通讯作者：
Holmes,EW
中科院分区：
文献类型：
--
作者：
Ch'ng,JL;Mulligan,RC;Schimmel,P;Holmes,EW
反义RNA是产生显性失活突变的潜在强大工具，但该策略的局限性之一是反义转录物在阻断靶基因表达方面相对低效。为了识别更有效的靶序列，使用无辅助逆转录病毒介导的基因转移将与人肌酸激酶B (CK-B) mRNA的多个功能区互补的反义RNA引入U937细胞中。与该 mRNA 的最后三分之一编码区和所有非编码区互补的反义 RNA 非常有效；一两个反义转录物足以阻断一种 CK-B mRNA 的表达。相反，与最后17个密码子互补的序列和所有3'非编码区已被删除的反义RNA对CK-B表达没有影响。反义 RNA 都不会改变目标信息的丰度、初级转录物的处理、CK-B 信息从细胞核的输出或蔗糖梯度中 CK-B mRNA 的多核糖体谱。这些结果指出反义转录物对翻译的直接影响，并表明这种影响至少部分可以通过由于远端编码和/或3'非编码区中形成的双链体而抑制延伸或终止来解释。
Antisense RNA is a potentially powerful tool for creating dominant negative mutations, but one of the limitations of this strategy has been the relative inefficiency of antisense transcripts in blocking target gene expression. To identify more effective target sequences, helper-free retrovirus-mediated gene transfer was used to introduce antisense RNAs complementary to multiple functional regions of the human creatine kinase B (CK-B) mRNA into U937 cells. Antisense RNA complementary to the last third of the coding and all of the noncoding regio of this mRNA is highly effective; one or two antisense transcripts is sufficient to block the expression of one CK-B mRNA. In contrast, antisense RNA from which sequences complementary to the last 17 codons and all the 3' noncoding region have been deleted has no effect on CK-B expression. Neither antisense RNA alters the abundance of the target message, processing of the primary transcript, egress of the CK-B message from the nucleus, or the polysome profile of CK-B mRNA in sucrose gradients. These results point to a direct effect of the antisense transcript on translation and suggest that this effect may be explained at least in part by an inhibition of elongation or termination as a consequence of the duplex formed in the distal coding and/or 3' noncoding region.