CRISPR/Cas9-mediated ebony knockout results in puparium melanism in Spodoptera litura

• 来源期刊: 中国昆虫科学（英文版）
• 关键词: CRISPR/Cas9 ebony marker gene Spodoptera litura
• 年，卷（期）: 2019,（6）
• 页码范围: 1011-1019
• 页数: 9页
• 语种: 英文
• DOI: 10.1111/1744-7917.12663

摘要

Insect body pigmentation and coloration are critical to adaption to the environment.To explore the mechanisms that drive pigmentation,we used the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) genome editing system to target the ebony gene in the non-model insect Spodoptera litura.Ebony is crucial to melanin synthesis in insects.By directly injecting Cas9 messenger RNA and ebony-specific guide RNAs into S.litura embryos,we successfully induced a typical ebony-deficient phenotype of deep coloration of the puparium and induction of melanin formation during the pupal stage.Polymerase chain reaction-based genotype analysis demonstrated that various mutations had occurred at the sites targeted in ebony.Our study clearly demonstrates the function of ebony in the puparium coloration and also provides a potentially useful marker gene for functional studies in S.litura as well as other lepidopteran pests.