Department of Molecular Biology, The University of Texas Southwestern Medical Center, Dallas, TX, USA.
Methods Mol Biol. 2023;2677:99-112. doi: 10.1007/978-1-0716-3259-8_5.
CRISPR-Cas9 genome editing technology can be used to manipulate the genome of Drosophila melanogaster. The ability to delete genes, make specific mutations, add tags, or make other genetic manipulations is useful for studying germline stem cell biology. In this chapter, we will describe a method to use CRISPR-Cas9 genome editing technology to make knock-out and knock-in flies. We will cover everything from guideRNA (gRNA) and donor plasmid design and cloning to screening for positive edits.
CRISPR-Cas9 基因组编辑技术可用于操纵黑腹果蝇的基因组。删除基因、产生特定突变、添加标签或进行其他遗传操作的能力对于研究生殖干细胞生物学非常有用。在本章中，我们将描述一种使用 CRISPR-Cas9 基因组编辑技术制造敲除和敲入蝇的方法。我们将涵盖从引导 RNA（gRNA）和供体质粒设计和克隆到筛选阳性编辑的所有内容。
