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- W2775757142 abstract "Top of pageAbstractEpigenetic programming is mitotically heritable and affects gene expression without altering DNA sequence. The transition from active to inactive gene expression is referred to as gene silencing. Transgenes are often subject to partial gene silencing known as variegation. Variegation is highly dynamic in embryonic stem cells (ESC) with frequent switching between expressing and silent states.To examine variegation of a retrovirus vector in real time, a novel cell culture apparatus was used for long term imaging and eGFP transgene expression tracking over 4 days. A high resolution time course DIC and fluorescence image time sequence was obtained that examines variegation in expression levels of an MSCV-PGK-eGFP retrovirus vector in a mouse ESC colony. With this approach we recorded eGFP transgene expression levels from seven founder ESCs and their progeny over seven generations.We report that the variegated ESC line contained lineages that were stably off or stably on, indicating that their epigenetic program was stable over many generations. At the same time, other genetically identical lineages switched through an unstable but symmetrically inherited epigenetic program that initially produced similarly expressing or silenced progeny. Nevertheless, changes in expression patterns accumulate over subsequent generations indicating that the epigenetic program in these unstable lineages continues to change over time. In addition, dramatic asymmetric reprogramming can occasionally occur between siblings. To our knowledge, this is the first real time tracking of transgene variegation in mammalian cells and reveals a surprising diversity of epigenetic states. Our novel approach documents the dynamics and stability of retrovirus variegation in stem cells, and may facilitate efforts to overcome variegation through vector redesign or drug intervention." @default.
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- W2775757142 date "2005-05-01" @default.
- W2775757142 modified "2023-09-26" @default.
- W2775757142 title "79. Real Time Fluorescent Tracking of Dynamic Variegation by Retrovirus Vectors in Stem Cells" @default.
- W2775757142 doi "https://doi.org/10.1016/j.ymthe.2005.06.057" @default.
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