PDF(Pubmed)
Abstract：
The generation of knockins is fundamental to dissect biological systems. SEED/Harvest, a technology based on CRISPR-Cas9, offers a powerful approach for seamless genome editing in Drosophila. Here, we present a protocol to tag any gene in the Drosophila genome using SEED/Harvest technology. We describe knockin design, plasmid preparation, injection, and insertion screening. We then detail procedures for germline harvesting. The technique combines straightforward cloning and robust screening of insertions, while still resulting in scarless gene editing. For complete details on the use and execution of this protocol, please refer to Aguilar et al.1.
摘要：
敲除蛋白的产生是解剖生物系统的基础。种子/收获,基于CRISPR-Cas9的技术为果蝇的无缝基因组编辑提供了一种强大的方法。这里,我们提出了使用SEED/Harvest技术标记果蝇基因组中的任何基因的方案。我们描述了门金设计，质粒制备，注射,和插入筛查。然后，我们详细说明了种系收获的程序。该技术结合了简单的克隆和插入的强大筛选，同时仍然导致无疤痕的基因编辑。有关此协议的使用和执行的完整详细信息，请参考Aguilar等人1。
