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Published on: February 22, 2018

dsRNase1对Sf9细胞条件介质中dsRNA降解活性的贡献

Jinmo Koo¹, Subba Reddy Palli¹

¹Department of Entomology, College of Agriculture, University of Kentucky, Lexington, KY, United States.

Frontiers in insect science

|March 12, 2024

概括

科学家们确定了dsRNase1作为一种关键的酶,在Spodoptera frugiperda (Sf9) 细胞中降解双链RNA (dsRNA). 降低dsRNase1活动提高了这些昆虫的RNA干扰 (RNAi) 效率.

科学领域:

昆虫学昆虫学是一门学科.
分子生物学分子生物学
生物化学生物化学

背景情况:

在像Spodoptera frugiperda.这样的类昆虫中,RNA干扰 (RNAi) 是无效的.
在昆虫的血球和光线中高RNase活性有助于RNAi效率低.
Sf9细胞条件介质表现出显著的dsRNA降解活性,但负责的酶仍未确定.

研究的目的:

为了识别在Spodoptera frugiperda (Sf9) 细胞条件介质中对dSRNA降解负责的酶.
研究已识别的核酶在dsRNA降解和RNAi效率中的作用.
开发一种用于识别参与dSRNA降解的其他基因的系统.

主要方法:

在Sf9细胞中高表达核酶基因 (REase,RNaseT2,dsRNase1) 的表达分析.
用RNAi技术对候选核酶进行基因敲除.
从Sf9细胞的条件介质中测定dsRNA降解活性.
在核酶敲击后测量RNAi效率.

主要成果:

核酶基因REase,RNaseT2和dsRNase1被确定在Sf9细胞中高度表达.
在Sf9细胞条件介质中,dsrNase1的抑制显著降低了dsrna降解活性.
抑制dSRNase1导致Sf9细胞RNAi效率显著增加.

结论:

dsRNase1被确定为Sf9细胞条件介质中dsRNA降解活性的主要贡献者.
降低dsRNase1活动可以增强Spodoptera frugiperda中的RNAi效率.
描述了在Sf9细胞中识别其他dSRNA降解酶和RNAi调节剂的有效系统.

关键词:

它们是RNAiRNAi.它们是花鸟 (Spodoptera frugiperda) 的种类.dsRNA 的稳定性秋天的军队虫核酶核酶是什么意思

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