CONSTITUTIVE AND CONDITIONAL SUPPRESSION OF EXOGENOUS AND ENDOGENOUS GENES BY ANTI-SENSE RNA
CONSTITUTIVE AND CONDITIONAL SUPPRESSION OF EXOGENOUS AND ENDOGENOUS GENES BY ANTI-SENSE RNA
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DOI：
10.1126/science.2990048
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发表时间：
1985-01-01
期刊：
影响因子：
56.9
通讯作者：
WEINTRAUB, H
中科院分区：
文献类型：
--
作者：
IZANT, JG;WEINTRAUB, H
指导非编码（反义）DNA链转录的质粒DNA可以特异性抑制几种测试基因以及正常内源基因的表达。反义质粒构建体可以通过转染或显微注射引入真核细胞中，并在瞬时和稳定转化测定中发挥作用。与5′非翻译靶基因mRNA的52个碱基互补的反义转录物特异性抑制基因活性，与单独针对蛋白质编码结构域的反义转录物一样，或比单独针对蛋白质编码结构域的反义转录物更有效。条件性反义抑制通过使用酶诱导型启动子序列来实现。通过反义RNA对内源性肌动蛋白基因活性的抑制被检测为生长速率的降低和肌动蛋白微丝电缆的数量的减少。这些观察结果表明，反义RNA通常可用于抑制体内特定基因的表达，并且可能是经典遗传分析的潜在分子替代方案。
Plasmid DNA directing transcription of the noncoding (anti-sense) DNA strand can specifically inhibit the expression of several test genes as well as normal, endogenous genes. The anti-sense plasmid constructions can be introduced into eukaryotic cells by transfection or microinjection and function in both transient and stable transformation assays. Anti-sense transcripts complementary to as little as 52 bases of 5′ untranslated target gene mRNA specifically suppress gene activity as well as, or more efficiently than, anti-sense transcripts directed against the protein coding domain alone. Conditional anti-sense inhibition is accomplished with the use of hormone-inducible promoter sequences. Suppression of endogenous actin gene activity by anti-sense RNA is detected as a decrease in growth rate and as a reduction in the number of actin microfilament cables. These observations suggest that anti-sense RNA may be generally useful for suppressing the expression of specific genes in vivo and may be a potential molecular alternative to classical genetic analysis.