CRISPR 介导的埃及伊蚊基因组工程。
CRISPR-Mediated Genome Engineering in Aedes aegypti.
机构信息
Division of Biological Sciences, Section of Cell and Developmental Biology, University of California San Diego, La Jolla, CA, USA.
W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins Malaria Research Institute, Johns Hopkins Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, USA.
出版信息
Methods Mol Biol. 2022;2509:23-51. doi: 10.1007/978-1-0716-2380-0_2.
CRISPR-mediated genome engineering technologies have been adapted to a wide variety of organisms with high efficiency and specificity. The yellow fever mosquito, Aedes aegypti , is one such organism. It is also responsible for transmitting a wide variety of deadly viruses including Dengue, Zika, Yellow fever, and Chikungunya. The key to successful CRISPR-mediated gene editing applications is the delivery of both Cas9 ribonuclease and single-guide RNA (sgRNA ) to the nucleus of desired cells. Various methods have been developed for supplying the Cas9 endonuclease, sgRNA , and donor DNA to Ae. aegypti. In this chapter, we focus on methods of direct embryo delivery of editing components, presenting detailed step-by-step CRISPR/Cas9-based genome-editing protocols for inducing desired heritable edits in mosquitoes as well as insights into successful application of these protocols. We also highlight potential opportunities for customizing these protocols to manipulate the mosquito genome for innovative in vivo gene function studies.
CRISPR 介导的基因组编辑技术已经被高效和特异地应用于各种生物体。黄热病蚊子，埃及伊蚊，就是这样一种生物体。它还负责传播包括登革热、寨卡、黄热病和基孔肯雅热在内的多种致命病毒。成功的 CRISPR 介导的基因编辑应用的关键是将 Cas9 核糖核酸酶和单指导 RNA（sgRNA）递送到所需细胞的细胞核中。已经开发了各种方法来提供 Cas9 内切酶、sgRNA 和供体 DNA 给埃及伊蚊。在本章中，我们专注于直接胚胎递送编辑组件的方法，提供了详细的基于 CRISPR/Cas9 的基因组编辑方案，用于诱导蚊子中所需的可遗传编辑，以及成功应用这些方案的见解。我们还强调了定制这些方案以操纵蚊子基因组进行创新的体内基因功能研究的潜在机会。
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