Viable RNaseH1 knockout mice show RNaseH1 is essential for R loop processing, mitochondrial and liver function

被引:87
作者
Lima, Walt F. [1 ]
Murray, Heather M. [1 ]
Damle, Sagar S. [2 ]
Hart, Christopher E. [2 ]
Hung, Gene [3 ]
De Hoyos, Cheryl Li [1 ]
Liang, Xue-Hai [1 ]
Crooke, Stanley T. [1 ]
机构
[1] Ionis Pharmaceut Inc, Dept Core Antisense Res, 2855 Gazelle Court, Carlsbad, CA 92010 USA
[2] Ionis Pharmaceut Inc, Dept Funct Genom, 2855 Gazelle Court, Carlsbad, CA 92010 USA
[3] Ionis Pharmaceut Inc, Dept Antisense Drug Discovery, 2855 Gazelle Court, Carlsbad, CA 92010 USA
关键词
THERMUS-THERMOPHILUS HB8; RIBONUCLEASE-H; ESCHERICHIA-COLI; SACCHAROMYCES-CEREVISIAE; MOLECULAR-CLONING; DNA; PURIFICATION; SUBSTRATE; RNA; EXPRESSION;
D O I
10.1093/nar/gkw350
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Viable constitutive and tamoxifen inducible liver-specific RNase H1 knockout mice that expressed no RNase H1 activity in hepatocytes showed increased R-loop levels and reduced mitochondrial encoded DNA and mRNA levels, suggesting impaired mitochondrial R-loop processing, transcription and mitochondrial DNA replication. These changes resulted in mitochondrial dysfunction with marked changes in mitochondrial fusion, fission, morphology and transcriptional changes reflective of mitochondrial damage and stress. Liver degeneration ensued, as indicated by apoptosis, fibrosis and increased transaminase levels. Antisense oligonucleotides (ASOs) designed to serve as substrates for RNase H1 were inactive in the hepatocytes from the RNase H1 knockout mice and in vivo, demonstrating that RNase H1 is necessary for the activity of DNA-like ASOs. During liver regeneration, a clone of hepatocytes that expressed RNase H1 developed and partially restored mitochondrial and liver function.
引用
收藏
页码:5299 / 5312
页数:14
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