Gaul Connor R, Vijay Trisha, Johnson Rebecca, Macias Vanessa M
Department of Biological Sciences, University of North Texas, Denton, TX, USA.
Advanced Environmental Research Institute, University of North Texas, Denton, TX, USA.
Methods Mol Biol. 2025;2935:335-384. doi: 10.1007/978-1-0716-4583-3_15.
Site-specific genome editing is the most direct way to test gene function. When CRISPR-Cas9 was introduced for the editing of eukaryotic genomes, entomologists were ready with questions but had many methodologies to forge for the approach to be useful. Now, roughly 45 non-model insect genomes have been edited to study processes such as insecticide resistance, olfaction, immunity, and development. A useful first step for gene editing in an insect species of interest is identification and targeted editing of a gene with a visible phenotype. Visible markers increase the efficiency of detection of a genetic change; a wide availability of markers is one reason why model insects are so easy to manipulate and so have been key in understanding many biological processes. Here we will describe with detailed protocols how to approach a new insect species with CRISPR-era approaches by targeting a visual marker with Cas9-editing.
位点特异性基因组编辑是测试基因功能最直接的方法。当CRISPR-Cas9被引入用于真核生物基因组编辑时，昆虫学家虽有诸多疑问，但也有许多方法来使该方法变得有用。如今，大约45种非模式昆虫基因组已被编辑，用于研究诸如抗药性、嗅觉、免疫和发育等过程。在感兴趣的昆虫物种中进行基因编辑的一个有用的第一步是鉴定具有可见表型的基因并对其进行靶向编辑。可见标记提高了检测基因变化的效率；标记的广泛可用性是模式昆虫如此易于操作且在理解许多生物学过程中起关键作用的一个原因。在此，我们将通过详细的方案描述如何采用CRISPR时代的方法，通过用Cas9编辑靶向视觉标记来研究一种新的昆虫物种。
