CRISPR/Cas9-based knockout reveals that the clock genetimelessis indispensable for regulating circadian behavioral rhythms inBombyx mori
CRISPR/Cas9-based knockout reveals that the clock genetimelessis indispensable for regulating circadian behavioral rhythms inBombyx mori
复制标题
基于 CRISPR/Cas9 的敲除揭示了生物钟对于调节家蚕昼夜节律是不可或缺的
DOI：
10.1111/1744-7917.12864
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发表时间：
2020-09-18
期刊：
影响因子：
4
通讯作者：
Li, Mu-Wang
中科院分区：
文献类型：
--
作者：
Nartey, Moses Addo;Sun, Xia;Li, Mu-Wang
昼夜节律是普遍存在和适应的，存在于从微生物到人类的所有物种中，它们组织和改变行为和生理。无时(TIM)是一个典型的时钟基因。果蝇黑激素内源性生物钟的核心成分已被广泛研究；然而，在鳞翅目昆虫中，包括家蚕在内，其机制复杂，对Time参与负责行为活动的负反馈环路知之甚少。对时间的作用有一个全面的理解。我们开发了簇状规则间隔短回文重复序列(CRISPR)/CRISPR相关蛋白9基因编辑系统。我们试图阐明TIME在生物钟OB中的功能。近在咫尺的Bmtimmuters。基因敲除影响了两种昼夜行为活动：成虫羽化和胚胎孵化节律。实时定量聚合酶链式反应结果证实，TIM基因敲除导致Bmper和Bmclk信使RNA在光相和暗期的表达水平相对降低，从而导致振荡幅度降低。此外，在atim基因敲除中，BmdBt的每日节律性表达在光期上调，在暗期下调。我们的研究表明，时间对b是不可或缺的。Moricircadian时钟，可能参与调节羽化和孵化的节律。
Circadian rhythms, which are ubiquitous and adaptive, occur across all species, from microbes to humans, in which they organize and modify behavior and physiology.timeless(tim) is a canonical clock gene. The core composition of theDrosophila melanogasterendogenous circadian clock has been extensively investigated; however, in lepidopteran insects, includingBombyx mori, the mechanism is complicated and little is known regarding the participation oftimin the negative feedback loop responsible for behavioral activities. To arrive at a comprehensive understanding of the role oftimin theB. moriendogenous circadian clock, we exploited the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 gene editing system. We attempted to elucidate the functions oftimin the circadian clock ofB. moriusingBmtimmutants. The knockouts affected two circadian behavioral activities: adult emergence and embryo hatching rhythms. Quantitative real-time polymerase chain reaction results confirmed thattim-knockouts induced relative reductions in the expression levels, and thereby the oscillation amplitudes, ofBmperandBmclkmessenger RNAs during both the photophase and scotophase. Additionally, the daily rhythmic expression ofBmdbtwas upregulated in the photophase and downregulated in the scotophase in atim-knockout. Our study reveals thattimis integral to theB. moricircadian clock and may be involved in regulating eclosion and hatching rhythms.