Graduate Group in Biophysics, University of California San Francisco, San Francisco, CA, 94158, USA.
UC Berkeley - UCSF Graduate Program in Bioengineering, University of California San Francisco, San Francisco, CA, 94158, USA.
Nat Commun. 2019 May 9;10(1):2127. doi: 10.1038/s41467-019-09985-2.
The CRISPR-Cas9 system provides the ability to edit, repress, activate, or mark any gene (or DNA element) by pairing of a programmable single guide RNA (sgRNA) with a complementary sequence on the DNA target. Here we present a new method for small-molecule control of CRISPR-Cas9 function through insertion of RNA aptamers into the sgRNA. We show that CRISPR-Cas9-based gene repression (CRISPRi) can be either activated or deactivated in a dose-dependent fashion over a >10-fold dynamic range in response to two different small-molecule ligands. Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.
CRISPR-Cas9 系统提供了通过将可编程的单指导 RNA（sgRNA）与 DNA 靶标上的互补序列配对来编辑、抑制、激活或标记任何基因（或 DNA 元件）的能力。在这里，我们通过将 RNA 适体插入 sgRNA 中，提出了一种用于小分子控制 CRISPR-Cas9 功能的新方法。我们表明，基于 CRISPR-Cas9 的基因抑制（CRISPRi）可以在响应两种不同的小分子配体时以剂量依赖性方式被激活或失活，动态范围超过 10 倍。由于我们的系统直接作用于每个靶标特异性 sgRNA，因此它能够实现新的应用，这些应用需要对多个基因进行差异化和相反的时间控制。
