Retrotransposon Gene Engineering

We have used a mobile mouse VL30 genetic element together with retroviral helper cells to efficiently transmit and express chimeric foreign gene sequences in murine and human cells. The construct comprised a cDNA copy of retrotransposon NVL3, an internal promoter [rat cytosolic phosphoenolpyruvate c...

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Bibliographic Details
Published in:Bio/Technology 1991-08, Vol.9 (8), p.748-751
Main Authors: Cook, R. Frank, Cook, Sheila J, Hodgson, Clague P
Format: Article
Language:English
Subjects:
Agriculture
Animals
Base Sequence
Bioinformatics
Biological and medical sciences
Biomedical and Life Sciences
Biomedical Engineering/Biotechnology
Biomedicine
Biotechnology
Cell Line
Chimera
DNA Transposable Elements - genetics
DNA, Recombinant
Fundamental and applied biological sciences. Psychology
Genetic Vectors
Genic rearrangement. Recombination. Transposable element
Humans
Life Sciences
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
research-paper
Retroviridae - genetics
RNA-Directed DNA Polymerase - metabolism
Transduction, Genetic
Tumor Cells, Cultured
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Summary:We have used a mobile mouse VL30 genetic element together with retroviral helper cells to efficiently transmit and express chimeric foreign gene sequences in murine and human cells. The construct comprised a cDNA copy of retrotransposon NVL3, an internal promoter [rat cytosolic phosphoenolpyruvate carboxykinase (PEPCK, EC 4.1.1.32)] and an expressed bacterial neomycin resistance gene. Thirty to sixty thousand colony forming units/ml (CFU/ml) were recovered from the supernatant of mass cultured psi2 helper cells transfected with the recombinant retro-transposon plasmid DNA. RNA was expressed from both the VL30 long terminal repeat and from the internal PEPCK promoter, resulting in a G418 drug resistance phenotype in recipient cells. Integrated VL30 DNA sequences transduced from psi2 or PAS 17 retroviral helper cells failed to regenerate detectable replication competent virus. Human and rodent recipient cells transduced by the retrotransposons appeared to bear intact vector sequences after two rounds of transmission by helper cells.
ISSN:0733-222X
1087-0156
2331-3684
1546-1696
DOI:10.1038/nbt0891-748