Endogenous ADAR-mediated RNA editing in non-human primates using stereopure chemically modified oligonucleotides
Endogenous ADAR-mediated RNA editing in non-human primates using stereopure chemically modified oligonucleotides
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DOI：
10.1038/s41587-022-01225-1
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发表时间：
2022-03-07
影响因子：
46.9
通讯作者：
Vargeese, Chandra
中科院分区：
文献类型：
--
作者：
Monian, Prashant;Shivalila, Chikdu;Vargeese, Chandra
招募内源性RNA编辑酶并将其活性引导至特定细胞RNA的技术具有治疗潜力，但将其从细胞培养物转化为动物模型一直具有挑战性。在这里，我们描述了短的，化学修饰的寡核苷酸称为AIMers，直接有效和特异性的A到I编辑的内源性转录物的内源性腺苷脱氨酶作用于RNA（阿达尔）酶，包括无处不在的和组成型表达的ADAR 1 p110亚型。我们表明，完全化学修饰的AIMers与嵌合骨架含有立体纯硫代磷酸酯和含氮键的基础上磷酰胍增强的效力和编辑效率100倍，与那些均匀的硫代磷酸酯修饰的骨架在体外。在体内，用N-乙酰半乳糖胺靶向肝细胞的AIM实现高达50%的编辑，而在非人灵长类动物肝脏中内源性ACTB转录物没有旁观者编辑，编辑持续至少一个月。这些结果支持进一步研究立体纯AIM的治疗潜力。使用短立体纯寡核苷酸有效编辑非人灵长类动物肝脏中的RNA。
Technologies that recruit and direct the activity of endogenous RNA-editing enzymes to specific cellular RNAs have therapeutic potential, but translating them from cell culture into animal models has been challenging. Here we describe short, chemically modified oligonucleotides called AIMers that direct efficient and specific A-to-I editing of endogenous transcripts by endogenous adenosine deaminases acting on RNA (ADAR) enzymes, including the ubiquitously and constitutively expressed ADAR1 p110 isoform. We show that fully chemically modified AIMers with chimeric backbones containing stereopure phosphorothioate and nitrogen-containing linkages based on phosphoryl guanidine enhanced potency and editing efficiency 100-fold compared with those with uniformly phosphorothioate-modified backbones in vitro. In vivo, AIMers targeted to hepatocytes with N-acetylgalactosamine achieve up to 50% editing with no bystander editing of the endogenous ACTB transcript in non-human primate liver, with editing persisting for at least one month. These results support further investigation of the therapeutic potential of stereopure AIMers.RNA in non-human primate liver is efficiently edited using short stereopure oligonucleotides.