Journal ArticleDOI
Cell electroporation is a highly efficient method for introducing restriction endonucleases into cells
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TLDR
Cell electroporation is used with great success to permeabilize Chinese hamster ovary cells for the introduction of restriction enzymes, resulting in extremely high frequencies (greater than 90%) of aberrant metaphase cells and also a dramatic decrease in cell survival, as measured by subsequent colony formation.Abstract:
Restriction endonucleases that make either blunt- or cohesive-end DNA double-strand breaks can induce chromosome aberrations. We have used cell electroporation with great success to permeabilize Chinese hamster ovary cells for the introduction of restriction enzymes. The introduction of restriction enzymes by this method resulted in extremely high frequencies (> 90%) of aberrant metaphase cells and also a dramatic decrease in cell survival, as measured by subsequent colony formation. Cell electroporation by itself caused no increase in aberrant chromosomes and had only a slight effect on cell survival.read more
Citations
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DNA strand breaks: the DNA template alterations that trigger p53-dependent DNA damage response pathways.
TL;DR: It is concluded that DNA strand breaks were sufficient for initiating p53-dependent signal transduction after finding that introduction of nucleases into cells by electroporation stimulated rapid p53 protein elevations.
Journal ArticleDOI
Integration of DNA fragments by illegitimate recombination in Saccharomyces cerevisiae
TL;DR: Results indicates that four base pairs of homology are sufficient for some types of mitotic recombination.
Journal ArticleDOI
Cell electropermeabilization: a new tool for biochemical and pharmacological studies.
Stéphane Orlowski,Lluis M. Mir +1 more
TL;DR: Electropermeabilization of the plasma membrane by means of short and intense electric pulses allows pharmacological studies with cells, cultured monolayers and in vivo tissues as well as the design of drug controlled-release systems.
Journal ArticleDOI
Restriction enzyme-mediated DNA integration in Coprinus cinereus
TL;DR: This method is analysed in the basidiomycete Coprinus cinereus using the homologous pab1 gene as a selectable marker and the restriction enzymes BamHI, EcoRI and PstI to identify the enzyme concentration that gives the highest number of transformants in an enzyme- and concentration-dependent manner.
Journal ArticleDOI
Illegitimate recombination induced by DNA double-strand breaks in a mammalian chromosome.
J W Phillips,William F. Morgan +1 more
TL;DR: DNA double-strand-break-induced mutations in the endogenous adenine phosphoribosyl-transferase (APRT) gene in cultured Chinese hamster ovary cells after exposure to restriction endonucleases indicate that blunt-end DNA double- STRands can induce illegitimate (nonhomologous) recombination in mammalian chromosomes and that they play an important role in mutagenesis.
References
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Journal ArticleDOI
Enzymatic restriction of mammalian cell DNA using Pvu II and Bam H1: evidence for the double-strand break origin of chromosomal aberrations.
TL;DR: The method of introducing restriction enzymes into cells opens up a new model approach for the study of the conversion of double-strand breaks into chromosome aberrations, which may be easily repaired by the cell due to the stabilization of the hydrogen bonding at the site of the double- Strand break.
Journal ArticleDOI
Molecular mechanisms involved in the production of chromosomal aberrations. III: Restriction endonucleases
TL;DR: In this article, Chinese hamster ovary cells (CHO cells) and mouse fibroblasts (PG 19) were permeabilized with inactivated Sendai virus, treated with different types of restriction endonucleases (Eco RV, Pvu II, Bam HI, Sma I, Asu III, Nun II), and studied for the occurrence of chromosomal aberrations at different times following treatment.
Journal Article
Introduction and expression of DNA molecules in eukaryotic cells by electroporation.
G L Andreason,G A Evans +1 more
TL;DR: Electroporation and simple, transient expression assays using indicator genes allow the calibration and optimization of electroporation conditions for a wide variety of eukaryotic cell types and may allow the genetic modification of a variety of host cells which cannot be easily transformed by other methods.
Journal ArticleDOI
Enzymatic restriction of mammalian cell DNA: evidence for double-strand breaks as potentially lethal lesions.
TL;DR: The results reported here show that Pvu II simulates X-ray exposure, in causing a dose-dependent loss of the reproductive integrity of mammalian cells, and support the notion that X-irradiated mammalian cells undergo a mode of death in which dsb in the DNA cause chromosomal aberrations which are lethal as a result of loss of genetic material in the form of chromosome fragments, or as the result of chromosome bridge formation.
Journal ArticleDOI
Chromosomal aberrations induced by restriction endonucleases.
TL;DR: Restriction endonucleases (REs) are able to induce chromosomal aberrations in Chinese hamster ovary (CHO) cells and these capacities are probably correlated with the number of recognition sites in the genome.