Department of Radiation Oncology, Washington University School of Medicine, St. Louis, MO, USA.
Methods Mol Biol. 2020;2115:351-364. doi: 10.1007/978-1-0716-0290-4_19.
A critical stage in performing gene editing experiments using the CRISPR/Cas9 system is the design of guide RNA (gRNA). In this chapter, we conduct a review of the current gRNA design rules for maximizing on-target Cas9 activity while minimizing off-target activity. In addition, we present some of the currently available computational tools for gRNA activity prediction and assay design.
在使用 CRISPR/Cas9 系统进行基因编辑实验的关键阶段是指导 RNA（gRNA）的设计。在本章中，我们对当前的 gRNA 设计规则进行了综述，以最大限度地提高靶 Cas9 活性，同时最小化脱靶活性。此外，我们还介绍了一些目前可用的用于 gRNA 活性预测和检测设计的计算工具。
