Nganso Beatrice T, Pines Gur, Soroker Victoria
Department of Entomology, Chemistry and Nematology, Institute of Plant Protection, Agricultural Research Organization, The Volcani Centre, Rishon LeZion, Israel.
Insect Biochem Mol Biol. 2022 Apr;143:103705. doi: 10.1016/j.ibmb.2021.103705. Epub 2022 Feb 5.
Functional genomics is an essential tool for elucidating the structure and function of genes in any living organism. Here, we review the use of different gene manipulation techniques in functional genomics of Acari (mites and ticks). Some of these Acari species inflict severe economic losses to managed crops and health problems to humans, wild and domestic animals, but many also provide important ecosystem services worldwide. Currently, RNA interference (RNAi) is the leading gene expression manipulation tool followed by gene editing via the bacterial type II Clustered Regularly Interspaced Short Palindromic Repeats and associated protein 9 system (CRISPR-Cas9). Whilst RNAi, via siRNA, does not always lead to expected outcomes, the exploitations of the CRISPR systems in Acari are still in their infancy and are limited only to CRISP/Cas9 to date. In this review, we discuss the advantages and disadvantages of RNAi and CRISPR-Cas9 and the technical challenges associated with their exploitations. We also compare the biochemical machinery of RNAi and CRISPR-Cas9 technologies. We highlight some potential solutions for experimental optimization of each mechanism in gene function studies. The potential benefits of adopting various CRISPR-Cas9 systems for expanding on functional genomics experiments in Acari are also discussed.
功能基因组学是阐明任何生物体中基因结构和功能的重要工具。在此,我们综述了不同基因操作技术在螨类(螨虫和蜱虫)功能基因组学中的应用。其中一些螨类物种给农作物造成严重经济损失,并给人类、野生动物和家畜带来健康问题,但许多螨类在全球范围内也提供重要的生态系统服务。目前,RNA干扰(RNAi)是主要的基因表达操作工具,其次是通过细菌II型成簇规律间隔短回文重复序列及其相关蛋白9系统(CRISPR-Cas9)进行基因编辑。虽然通过小干扰RNA(siRNA)的RNAi并不总是能产生预期结果,但CRISPR系统在螨类中的应用仍处于起步阶段,迄今为止仅限于CRISPR/Cas9。在本综述中,我们讨论了RNAi和CRISPR-Cas9的优缺点以及与其应用相关的技术挑战。我们还比较了RNAi和CRISPR-Cas9技术的生化机制。我们强调了在基因功能研究中对每种机制进行实验优化的一些潜在解决方案。还讨论了采用各种CRISPR-Cas9系统扩展螨类功能基因组学实验的潜在益处。