Aptazyme-embedded guide RNAs enable ligand-responsive genome editing and transcriptional activation.
Aptazyme-embedded guide RNAs enable ligand-responsive genome editing and transcriptional activation.
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DOI：
10.1038/ncomms15939
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发表时间：
2017-06-28
影响因子：
16.6
通讯作者：
Liu DR
中科院分区：
文献类型：
--
作者：
Tang W;Hu JH;Liu DR
可编程序列特异性基因组编辑剂（例如 CRISPR-Cas9）极大地提高了我们操纵人类基因组的能力。尽管基因组编辑剂和可编程转录调节剂的规范形式具有组成性活性，但对基因组编辑和转录调节活动的精确时间和空间控制将使这些强大技术的使用更有选择性且可能更安全。在这里，通过将配体响应性自切割催化RNA（适体酶）整合到引导RNA中，我们开发了一组嵌入适体酶的引导RNA，它们能够实现小分子控制的核酸酶介导的基因组编辑和小分子控制的碱基编辑，以及哺乳动物细胞中小分子依赖性转录激活。 CRISPR-Cas9 是一种用于操纵人类基因组的强大技术，但对活性的时间和空间控制将有助于更安全、更有选择性的使用。在这里，作者将适体酶整合到引导 RNA 中，以允许小分子激活 CRISPR-Cas9。
Programmable sequence-specific genome editing agents such as CRISPR-Cas9 have greatly advanced our ability to manipulate the human genome. Although canonical forms of genome-editing agents and programmable transcriptional regulators are constitutively active, precise temporal and spatial control over genome editing and transcriptional regulation activities would enable the more selective and potentially safer use of these powerful technologies. Here, by incorporating ligand-responsive self-cleaving catalytic RNAs (aptazymes) into guide RNAs, we developed a set of aptazyme-embedded guide RNAs that enable small molecule-controlled nuclease-mediated genome editing and small molecule-controlled base editing, as well as small molecule-dependent transcriptional activation in mammalian cells. CRISPR-Cas9 is a powerful technique for manipulating the human genome, however temporal and spatial control of activity would facilitate safer, more selective use. Here the authors incorporate aptazymes into guide RNAs to allow small molecule activation of CRISPR-Cas9.