Construction of mouse embryonic stem cells for the detection of gene mutation

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The mouse ES cells can be maintained in tissue culture with pluripotency. ES cells can also be induced to differentiate in vitro. These properties make it an excellent tool for the study of early embryogenesis and, potentially, teratogenesis in vitro. We have established genetic reporter systems in the ES cells for the detection of mutations. E.coli xanthine-guanine phosporibosyl transferase (XPRT) gene (gpt) was transfected to the HPRT- ES cells. After transfection with the gpt gene (pSV2-gpt), surviving cells producing XPRT can be selectively grown with xanthine as the sole precursor for guanine nucleotide formation in a medium containing inhibitors (aminopterin and mycophenolic acid) that block de novo purinenucleotide synthesis. After methyl methane sulfonate (MMS) exposure, signifcant increases of 6-TG resistant colony were observed.