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Stable reduction of thymidine kinase activity in cells expressing high levels of anti-sense RNA

Kim, Stuart K. and Wold, Barbara J. (1985) Stable reduction of thymidine kinase activity in cells expressing high levels of anti-sense RNA. Cell, 42 (1). pp. 129-138. ISSN 0092-8674. https://resolver.caltech.edu/CaltechAUTHORS:20190508-081413730

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Abstract

Anti-sense thymidine kinase (TK) RNA was expressed as part of a chimeric dihydrofolate reductase (DHFR) anti-sense TK transcript. High level expression was obtained by selection of cells resistant to progressively higher levels of methotrexate, provided by over-production of DHFR. The result was a concomitant increase in intracellular anti-sense TK RNA level owing to its presence on the same transcription unit as DHFR. In several cell lines expressing high levels of anti-sense TK RNA, thymidine kinase activity was reduced by 80%–90%. RNA:RNA duplexes were detected in the nuclear fraction. The results suggest a mechanism for diminution of TK activity; anti-sense RNA hybridizes with sense TK RNA in the nucleus, and duplex containing TK transcripts fail to enter the cytoplasm with normal efficiency.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/S0092-8674(85)80108-2DOIArticle
ORCID:
AuthorORCID
Wold, Barbara J.0000-0003-3235-8130
Additional Information:© 1985 by MIT. Received April 16, 1985; revised May 9, 1985. The authors thank Dr. Stewart Scherer and Paul Mueller for interesting discussions and suggestions during the course of this work, and on the manuscript. We thank Drs. Hal Weintraub and John Izant for interesting and enthusiastic conversations regarding anti-sense RNA. We also thank Professors Norman Davidson, Eric Davidson, and Joan Steitz and Anne Villenueve for critically reading the manuscript. This work was supported by a National Institutes of Health predoctoral training award to S. K. K., and the National Institutes of Health grant HL28021 to B. W. B. W. is a Rita Allen Foundation Fellow and a Searle Scholar, and is grateful for their support. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
Funders:
Funding AgencyGrant Number
NIH Predoctoral FellowshipUNSPECIFIED
NIHHL28021
Rita Allen FoundationUNSPECIFIED
Searle Scholars ProgramUNSPECIFIED
Issue or Number:1
Record Number:CaltechAUTHORS:20190508-081413730
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20190508-081413730
Official Citation:Stuart K. Kim, Barbara J. Wold, Stable reduction of thymidine kinase activity in cells expressing high levels of anti-sense RNA, Cell, Volume 42, Issue 1, 1985, Pages 129-138, ISSN 0092-8674, https://doi.org/10.1016/S0092-8674(85)80108-2. (http://www.sciencedirect.com/science/article/pii/S0092867485801082)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:95334
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:08 May 2019 15:54
Last Modified:03 Oct 2019 21:12

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