Department of Zoology, The University of British Columbia, Vancouver, British Columbia V6T 1Z4, Canada.
Department of Zoology, The University of British Columbia, Vancouver, British Columbia V6T 1Z4, Canada
Cold Spring Harb Protoc. 2024 Sep 3;2024(9):pdb.top107688. doi: 10.1101/pdb.top107688.
CRISPR-Cas9 has revolutionized gene editing for traditional and nontraditional model organisms alike. This tool has opened the door to new mechanistic studies of basic mosquito biology as well as the development of novel vector control strategies based on CRISPR-Cas9, including gene drives that spread genetic elements in the population. Although the promise of the specificity, flexibility, and ease of deployment CRISPR is real, its implementation still requires empirical optimization for each new species of interest, as well as to each genomic target within a given species. Here, we provide an overview of designing and testing single-guide RNAs for the use of CRISPR-based gene editing tools.
CRISPR-Cas9 技术已经彻底改变了传统和非传统模式生物的基因编辑。这项工具为基础蚊虫生物学的新机制研究以及基于 CRISPR-Cas9 的新型载体控制策略的发展打开了大门，包括在种群中传播遗传要素的基因驱动。尽管 CRISPR 的特异性、灵活性和易于部署的承诺是真实存在的，但它的实施仍然需要针对每个感兴趣的新物种以及给定物种内的每个基因组目标进行实证优化。在这里，我们提供了一个概述，用于设计和测试基于 CRISPR 的基因编辑工具的单指导 RNA。
