Podophyllotoxin resistance: a codominant selection system for quantitative mutagenesis studies in mammalian cells
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Abstract:
Mutants resistant to the microtubule inhibitor podophyllotoxin (PodR), a codominant marker, can be readily selected in various mammalian cell lines such as, CHO, HeLa, mouse L cells, Syrian hamster cells (BHK21) and a mouse teratocarcinoma cell line OC15. In CHO cells, the recovery of PodR mutants is not affected by cell density (up to 1 X 10(6) cells per 100-mm diameter dish), and after treatment with the mutagen ethyl methanesulfonate maximum mutagenic effect is achieved after a relatively short expression time (40-48 h). The frequency of PodR mutants in various cell lines increased in a dose-dependent manner in response to treatment with the mutagens ethyl methanesulfonate and N-methyl-N'-nitro-N-nitrosoguanidine. The PodR selection system thus provides a new genetic marker which should prove useful in studies of quantitative mutagenesis in mammalian cells.
Insights
Researchers developed a new genetic marker using podophyllotoxin-resistant (PodR) mutants for studying mammalian cell mutagenesis. This system allows for efficient selection of mutants across various cell lines, aiding quantitative mutagenesis research.
Area of Science:
- Cell Biology
- Genetics
- Toxicology
Background:
- Mammalian cell mutagenesis studies require reliable genetic markers.
- Microtubule inhibitors like podophyllotoxin offer potential for selection systems.
Purpose of the Study:
- To establish and characterize a novel genetic marker for quantitative mutagenesis in mammalian cells.
- To evaluate the utility of podophyllotoxin resistance (PodR) as a selectable marker.
Main Methods:
- Selection of podophyllotoxin-resistant (PodR) mutants in various mammalian cell lines (CHO, HeLa, mouse L, BHK21, OC15).
- Assessment of mutant recovery across different cell densities.
- Determination of optimal expression time for mutagenic effects.
- Dose-dependent analysis of mutant frequency following treatment with ethyl methanesulfonate and N-methyl-N'-nitro-N-nitrosoguanidine.
Main Results:
- PodR mutants were readily selected in multiple mammalian cell lines.
- Mutant recovery in CHO cells was independent of cell density.
- Optimal mutagenic effect in CHO cells occurred after 40-48 h expression time.
- Mutagen treatment resulted in a dose-dependent increase in PodR mutant frequency.
Conclusions:
- The podophyllotoxin resistance (PodR) system serves as a valuable new genetic marker.
- This system facilitates quantitative mutagenesis studies in mammalian cells.
- The PodR marker system demonstrates broad applicability across diverse cell lines.