Gene Expression from Transcriptionally Disabled Retroviral Vectors

Retroviral vectors are used for the efficient transfer of foreign genes into mammalian cells. We report here the construction of murine retrovirus-based vectors carrying the full-length cDNA for human hypoxanthine phosphoribosyltransferase (HPRT; EC 2.4.2.8) and from which the enhancer sequences, th...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1987-08, Vol.84 (15), p.5197-5201
Main Authors: Yee, Jiing-Kuan, Moores, Jane C., Jolly, Douglas J., Wolff, Jon A., Respess, James G., Friedmann, Theodore
Format: Article
Language:English
Subjects:
3T3 cells
Base Sequence
Biological and medical sciences
Biotechnology
Cell lines
Chromosome Deletion
cloning vectors
Complementary DNA
DNA
DNA - analysis
DNA, Recombinant - analysis
DNA, Viral - analysis
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation
gene transfer
Genes
Genetic engineering
Genetic technics
Genetic vectors
Humans
hypoxanthine phosphoribosyltransferase
Hypoxanthine Phosphoribosyltransferase - genetics
mammalian cells
Methods. Procedures. Technologies
Packaging
Repetitive Sequences, Nucleic Acid
Retroviridae - genetics
retrovirus
RNA
TATA box
Transcription, Genetic
Vectors (cloning, transfer, expression). Insertion sequences and transposons
Viruses
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Summary:Retroviral vectors are used for the efficient transfer of foreign genes into mammalian cells. We report here the construction of murine retrovirus-based vectors carrying the full-length cDNA for human hypoxanthine phosphoribosyltransferase (HPRT; EC 2.4.2.8) and from which the enhancer sequences, the ``CAAT box,'' and the ``TATA box'' in the long terminal repeats (LTRs) have been deleted. After infection of HPRT-deficient rat cells by the vectors, transcriptional activity from the 5′ LTR was undetectable and expression of the HPRT cDNA was dependent on an internal promoter. Removal of the LTR regulatory elements increased HPRT gene expression from an internal promoter, indicating interference between the two sets of transcriptional signals. Such disabled vectors may reduce the likelihood of undesirable genetic changes through insertional mutagenesis in cells infected with retroviral vectors.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.84.15.5197