RNase H1-Dependent Antisense Oligonucleotides Are Robustly Active in Directing RNA Cleavage in Both the Cytoplasm and the Nucleus.
RNase H1-Dependent Antisense Oligonucleotides Are Robustly Active in Directing RNA Cleavage in Both the Cytoplasm and the Nucleus.
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DOI：
10.1016/j.ymthe.2017.06.002
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发表时间：
2017-09-06
期刊：
影响因子：
--
通讯作者：
Crooke ST
中科院分区：
文献类型：
--
作者：
Liang XH;Sun H;Nichols JG;Crooke ST
依赖于RNaseH1的反义寡核苷酸(ASO)能有效地降低细胞质和核内滞留RNA的水平。虽然ASO在细胞核中的活性已被很好地证明，但ASO的细胞质活性尚不清楚。利用动力学和亚细胞分级研究，我们评估了细胞质中ASO的活性。在转染后，靶向外显子区域的ASO迅速减少细胞质中丰富的mRNAs，而仅降解核前mRNA的内含子靶向ASO降低mRNA水平的速度较慢，类似于正常的mRNA衰变。重要的是，一些外显子靶向ASO可以在不降低前-mRNA水平的情况下迅速而有力地降低mRNA水平，这表明预先存在的细胞质mRNAs可以被RNaseH1-ASO处理所切割。此外，我们还表达了一个细胞质定位的突变体7SL RNA，它含有部分U16小核仁RNA(SnoRNA)序列。ASO处理后，以RNaseH1依赖的方式，最早在30min就同时减少了核U16 snoRNA和细胞质7SL突变RNA。7SL突变体RNA的5‘和3’端裂解产物均在细胞质中积累。综上所述，这些结果表明依赖于RNaseH1的ASO在细胞质和细胞核中都非常活跃。梁等人。显示反义寡核苷酸(ASO)指导的RNaseH1裂解在细胞核和细胞质中都很活跃。他们发现ASOS可以触发由RNaseH1介导的细胞质和核RNA的快速切割，RNaseH1也存在于细胞质中。
RNase H1-dependent antisense oligonucleotides (ASOs) are active in reducing levels of both cytoplasmic mRNAs and nuclear retained RNAs. Although ASO activity in the nucleus has been well demonstrated, the cytoplasmic activity of ASOs is less clear. Using kinetic and subcellular fractionation studies, we evaluated ASO activity in the cytoplasm. Upon transfection, ASOs targeting exonic regions rapidly reduced cytoplasmically enriched mRNAs, whereas an intron-targeting ASO that only degrades the nuclear pre-mRNA reduced mRNA levels at a slower rate, similar to normal mRNA decay. Importantly, some exon-targeting ASOs can rapidly and vigorously reduce mRNA levels without decreasing pre-mRNA levels, suggesting that pre-existing cytoplasmic mRNAs can be cleaved by RNase H1-ASO treatment. In addition, we expressed a cytoplasm-localized mutant 7SL RNA that contains a partial U16 small nucleolar RNA (snoRNA) sequence. Treatment with an ASO simultaneously reduced both the nuclear U16 snoRNA and the cytoplasmic 7SL mutant RNA as early as 30 min after transfection in an RNase H1-dependent manner. Both the 5′ and 3′ cleavage products of the 7SL mutant RNA were accumulated in the cytoplasm. Together, these results demonstrate that RNase H1-dependent ASOs are robustly active in both the cytoplasm and nucleus. Liang et al. show that antisense oligonucleotide (ASO)-directed RNase H1 cleavage is robustly active in both the nucleus and cytoplasm. They found that ASOs can trigger rapid cleavage of both cytoplasmic and nuclear RNAs mediated by RNase H1, which is also present in the cytosol.
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DOI：
10.1038/nrg3355
发表时间：
2013-02
期刊：
Nature reviews. Genetics
影响因子：
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作者：
通讯作者：
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影响因子：
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作者：
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DOI：
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发表时间：
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影响因子：
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DOI：
10.1016/j.bbrc.2015.06.171
发表时间：
2015-08-21
影响因子：
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作者：
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