INHIBITION OF THYMIDINE KINASE GENE-EXPRESSION BY ANTI-SENSE RNA - A MOLECULAR APPROACH TO GENETIC-ANALYSIS
INHIBITION OF THYMIDINE KINASE GENE-EXPRESSION BY ANTI-SENSE RNA - A MOLECULAR APPROACH TO GENETIC-ANALYSIS
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DOI：
10.1016/0092-8674(84)90050-3
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发表时间：
1984-01-01
期刊：
影响因子：
64.5
通讯作者：
WEINTRAUB, H
中科院分区：
文献类型：
--
作者：
IZANT, JG;WEINTRAUB, H
As an alternative approach to classical genetic analysis, the potential of anti-sense (nonsense) DNA strand transcription to inhibit gene activity was investigated. A promoter will direct transcription of the complementary nonsense DNA strand when the protein coding sequence on a cloned gene is excised and then reinserted in reverse orientation. When such flipped gene constructions of the HSV [herpes simplex virus] thymidine kinase (TK) gene are coinjected with the wild-type gene at a 100:1 ratio, there is a reduction of transient TK expression in TK- mouse L cells. The proportion of viable cells with demonstrable TK activity drops 4-fold as compared with neighboring cells coinjected with TK and an excess of control plasmid. Autoradiography of the cells still expressing TK shows that 3H-thymidine incorporation is reduced. Cells cotransformed with flipped TK gene constructions have a reduced capacity to express subsequently microinjected TK genes, suggesting that the anti-message phenomenon is due to a trans-inhibition of TK and is probably not an artifact of rearrangements following microinjection.
