Widespread use of gene therapy technologies is limited in part by the lack of small genetic switches with wide dynamic ranges that control transgene expression without the requirement of additional protein components1-5. In this study, we engineered a class of type III hammerhead ribozymes to develop RNA switches that are highly efficient at cis-cleaving mammalian mRNAs and showed that they can be tightly regulated by a steric-blocking antisense oligonucleotide. Our variant ribozymes enabled in vivo regulation of adeno-associated virus (AAV)-delivered transgenes, allowing dose-dependent and up to 223-fold regulation of protein expression over at least 43 weeks. To test the potential of these reversible on-switches in gene therapy for anemia of chronic kidney disease6, we demonstrated regulated expression of physiological levels of erythropoietin with a well-tolerated dose of the inducer oligonucleotide. These small, modular and efficient RNA switches may improve the safety and efficacy of gene therapies and broaden their use.

中文翻译：

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可逆的RNA开关控制体内AAV传递的治疗药物的基因表达。

基因治疗技术的广泛使用部分受到缺乏小型遗传开关的限制，这些遗传开关具有宽动态范围，可控制转基因表达，而无需其他蛋白质组分1-5。在这项研究中，我们设计了一类III型锤头状核酶，以开发在顺式裂解哺乳动物mRNA上高效的RNA开关，并显示它们可以被空间封闭反义寡核苷酸严格调控。我们的变体核酶能够在体内调节腺相关病毒（AAV）传递的转基因，从而在至少43周内实现剂量依赖性和高达223倍的蛋白质表达调节。为了测试这些可逆接通开关在治疗慢性肾脏病贫血的基因治疗中的潜力6，我们证明了具有良好耐受剂量的诱导剂寡核苷酸可调节促红细胞生成素的生理水平表达。这些小型，模块化和高效的RNA开关可提高基因疗法的安全性和有效性，并扩大其用途。