Using CRISPR-Cas9 to create knockout mutants in insects

V. E. Hillary, S. A. Ceasar
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Abstract

Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (CRISPR/Cas9) is a potent genome editing tool that has rapidly revolutionized research in natural science disciplines. The CRISPR/Cas9 system surpasses traditional zinc finger nucleases and transcription activator-like effector nucleases because of its simplicity in design, cost-effectiveness, and labor intensity. The CRISPR/Cas9-mediated genome editing system has been used to generate precise and targeted mutations in many different organisms, including insects. However, there is still a lack of a comprehensive review that systematically summarizes the successful utilization of the CRISPR/Cas9 system in generating knockout (KO) mutations in insects (both model and non-model). Thus, this review provides a comprehensive analysis of the current advancements in CRISPR/Cas9-based KO mutants in insects, including both model and non-model species. We also discussed the limitations of the CRISPR/Cas9 system. Finally, we discuss the biosafety concerns associated with the release of CRISPR/Cas9-edited insects into the environment and draw inroads and insights into the future application of the CRISPR/Cas9 system in insects.
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利用 CRISPR-Cas9 在昆虫中制造基因敲除突变体
簇状规则间隔短回文重复序列(CRISPR)和 CRISPR 相关蛋白 9(CRISPR/Cas9)是一种有效的基因组编辑工具,它迅速给自然科学学科的研究带来了革命性的变化。CRISPR/Cas9 系统超越了传统的锌指核酸酶和转录激活剂样效应核酸酶,因为它设计简单、成本效益高、劳动强度低。CRISPR/Cas9 介导的基因组编辑系统已被用于在包括昆虫在内的许多不同生物体中产生精确的靶向突变。然而,目前仍缺乏系统总结 CRISPR/Cas9 系统成功用于在昆虫(包括模式和非模式)中产生基因敲除(KO)突变的全面综述。因此,本综述全面分析了目前基于CRISPR/Cas9的昆虫(包括模式和非模式物种)KO突变的进展。我们还讨论了 CRISPR/Cas9 系统的局限性。最后,我们讨论了将 CRISPR/Cas9 编辑过的昆虫释放到环境中的相关生物安全问题,并得出了 CRISPR/Cas9 系统未来在昆虫中应用的进展和见解。
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