Genomic instability in both wild-type and telomerase null MEFs
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- Hao, LY. & Greider, C.W. Chromosoma (2004) 113: 62. doi:10.1007/s00412-004-0291-7
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To examine chromosome instability in the absence of telomerase, we established mouse embryonic fibroblast (MEF) lines from late generation mTR−/− and wild-type animals and examined metaphases using telomere fluorescence in situ hybridization (FISH) and spectral karyotyping (SKY). In early passages, mTR−/− G6 cell lines showed more chromosome ends with no telomere signal, more chromosome end-to-end fusions and greater radiosensitivity than wild-type lines. At later passages, however, the rate of genomic instability in the wild-type MEFs increased to a level similar or higher than seen in the mTR−/− G6 cell lines. This high degree of instability in wild-type MEF lines suggests that post-crisis MEFs should not be considered genetically defined cell lines. Surprisingly, the increased radiosensitivity seen in early passage mTR−/− G6 cultures was lost after crisis. Both post-crisis mTR−/− G6 MEFs and wild-type MEFs showed loss of p53 and γ-H2AX phosphorylation in response to irradiation, indicating a loss of DNA damage checkpoints.