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Assays for compounds that modulate rhadino virus LANA action in trans on a unit of rhadino virus DNA to mediate efficient episome persistence
Assays for compounds that modulate rhadino virus LANA action in trans on a unit of rhadino virus DNA to mediate efficient episome persistence
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机译:调节狂犬病病毒DNA单位上的狂犬病病毒LANA反式调节介导有效附加体持久性的化合物的测定
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摘要
Primary effusion lymphoma (PEL) cells harbor Kaposi's sarcoma-associated herpesvirus (KSHV) episomes and express a KSHV encoded latency-associated nuclear antigen (LANA). In PEL cells, LANA and KSHV DNA co-localized in dots in interphase nuclei and along mitotic chromosomes. In the absence of KSHV DNA, LANA was diffusely distributed in the nucleus or on mitotic chromosomes. In lymphoblasts, LANA was necessary and sufficient for the persistence of episomes containing a specific KSHV DNA fragment. Furthermore, LANA co-localized with the artificial KSHV DNA episomes in nuclei and along mitotic chromosomes. The KSHV DNA segment that provides for efficient persistence in LANA positive cells has been identified as the rhodino virus cis-acting element (RVCAE). These results support a model in which LANA tethers episomes containing the KSHV RVCAE DNA to chromosomes during mitosis to enable efficient segregation to progeny cells. The products and methods of the invention are useful in identifying compounds for modulating (especially including interfering with) the persistance of rhodino virus DNA in mammalian cells, and disease states associated therewith, as well as such compounds themselves. The products and methods of the invention are also useful in modulating (especially including enabling or improving) the efficient persistence of a heterologous DNA containing RVCAE and a selected DNA in mammalian cells in which LANA is expressed for use in gene therapies and related techniques based on the selected DNA.
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