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• W1529866730 endingPage "592" @default.
• W1529866730 startingPage "583" @default.
• W1529866730 abstract "We have analyzed the response of a number of human cell lines to treatment with antisense oligodeoxynucleotides (ODNs) directed against RNA polymerase II, replication protein A, and Ha‐ ras . ODN‐delivery to the cells was liposome‐mediated or via electroporation, which resulted in different intracellular locations of the ODNs. The ODN‐mediated target mRNA reduction varied considerably between the cell lines. In view of the essential role of RNase H activity in this response, RNase H was analyzed. The mRNA levels of RNase H1 and RNase H2 varied considerably in the cell lines examined in this study. The intracellular localization of the enzymes, assayed by green‐fluorescent protein fusions, showed that RNase H1 was present throughout the whole cell for all cell types analyzed, whereas RNase H2 was restricted to the nucleus in all cells except the prostate cancer line 15PC3 that expressed the protein throughout the cell. Whole cell extracts of the cell lines yielded similar RNase H cleavage activity in an in vitro liquid assay, in contrast to the efficacy of the ODNs in vivo . Overexpression of RNase H2 did not affect the response to ODNs in vivo . Our data imply that in vivo RNase H activity is not only due to the activity assayed in vitro , but also to an intrinsic property of the cells. RNase H1 is not likely to be a major player in the antisense ODN‐mediated degradation of target mRNAs. RNase H2 is involved in the activity assayed in␣vitro . The presence of cell‐type specific factors affecting the activity and localization of RNase H2 is strongly suggested." @default.
• W1529866730 created "2016-06-24" @default.
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• W1529866730 date "2002-01-01" @default.
• W1529866730 modified "2023-10-17" @default.
• W1529866730 title "The involvement of human ribonucleases H1 and H2 in the variation of response of cells to antisense phosphorothioate oligonucleotides" @default.
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• W1529866730 doi "https://doi.org/10.1046/j.0014-2956.2001.02686.x" @default.
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