Fluorescent Guide RNAs Facilitate Development of Layered Pol II-Driven CRISPR Circuits,ACS Synthetic Biology

当前位置： X-MOL 学术 › ACS Synth. Biol. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Fluorescent Guide RNAs Facilitate Development of Layered Pol II-Driven CRISPR Circuits
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-07-18 00:00:00 , DOI: 10.1021/acssynbio.8b00153
David J Menn ₁ , Swechchha Pradhan ₁ , Samira Kiani ₁ , Xiao Wang ₁

Affiliation

Efficient clustered regularly interspaced short palindromic repeat (CRISPR) guide RNA (gRNA) expression from RNA Polymerase II (Pol II) promoters will aid in construction of complex CRISPR-based synthetic gene networks. Yet, we require tools to properly visualize gRNA directly to quantitatively study the corresponding network behavior. To address this need, we employed a fluorescent gRNA (fgRNA) to visualize synthetic CRISPR network dynamics without affecting gRNA functionality. We show that studying gRNA dynamics directly enables circuit modification and improvement of network function in Pol II-driven CRISPR circuits. This approach generates information necessary for optimizing the overall function of these networks and provides insight into the hurdles remaining in Pol II-regulated gRNA expression.

中文翻译：

荧光引导 RNA 促进分层 Pol II 驱动的 CRISPR 电路的开发

来自 RNA 聚合酶 II (Pol II) 启动子的高效成簇规则间隔短回文重复 (CRISPR) 引导 RNA (gRNA) 表达将有助于构建复杂的基于 CRISPR 的合成基因网络。然而，我们需要工具来正确地直接可视化 gRNA，以定量研究相应的网络行为。为了满足这一需求，我们采用荧光 gRNA (fgRNA) 来可视化合成 CRISPR 网络动态，而不影响 gRNA 功能。我们表明，研究 gRNA 动力学可以直接实现 Pol II 驱动的 CRISPR 电路中的电路修改和网络功能的改进。这种方法生成优化这些网络整体功能所需的信息，并提供对 Pol II 调节 gRNA 表达中剩余障碍的深入了解。

更新日期：2018-07-18

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11