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PMID: 2247070 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Expression of chimeric tRNA-driven antisense transcripts renders NIH 3T3 cells highly resistant to Moloney murine leukemia virus replication.

Molecular and cellular biology ·Vol. 10 ·No. 12 ·1990-12-00 ·Pages 6512-23

Sullenger BA, Lee TC, Smith CA, Ungers GE, Gilboa E

Abstract

NIH 3T3 cells infected with Moloney murine leukemia virus (MoMLV) express high levels of virus-specific RNA. To inhibit replication of the virus, we stably introduced chimeric tRNA genes encoding antisense templates into NIH 3T3 cells via a retroviral vector. Efficient expression of hybrid tRNA-MoMLV antisense transcripts and inhibition of MoMLV replication were dependent on the use of a particular type of retroviral vector, the double-copy vector, in which the chimeric tRNA gene was inserted in the 3' long terminal repeat. MoMLV replication was inhibited up to 97% in cells expressing antisense RNA corresponding to the gag gene and less than twofold in cells expressing antisense RNA corresponding to the pol gene. RNA and protein analyses suggest that inhibition was exerted at the level of translation. These results suggest that RNA polymerase III-based antisense inhibition systems can be used to inhibit highly expressed viral genes and render cells resistant to viral replication via intracellular immunization strategies.

MeSH Terms
Animals Cell Line Chimera Flow Cytometry Genetic Vectors Mice Moloney murine leukemia virus/genetics,physiology Protein Biosynthesis RNA, Antisense/genetics RNA, Transfer/genetics Templates, Genetic Transcription, Genetic Virus Replication
Chemicals
RNA, Antisense RNA, Transfer
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Sullenger B A
Program of Molecular Biology, Memorial Sloan-Kettering Cancer Center, New York, New York 10021.
Lee T C
Smith C A
Ungers G E
Gilboa E
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.10.12.6512-6523.1990
Published
1990-12-00
Pages
6512-23
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC362928
Subset
IM
Grants
NIAID NIH HHS · AI28771-02 · United States
External Links
PubMed source DOI: 10.1128/mcb.10.12.6512-6523.1990 PMC Full Text View journal details
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