RNA interference(RNAi)has been used for agricultural insect pest control based on silencing of targeted insect genes.However,the effectiveness of RNAi and its applications in insect pest control remain challenging.Her...RNA interference(RNAi)has been used for agricultural insect pest control based on silencing of targeted insect genes.However,the effectiveness of RNAi and its applications in insect pest control remain challenging.Here we review factors that may affect the effectiveness of RNAi application,including the variability in RNAi efficacy among different insect species,a limited understanding of double-stranded RNA(dsRNA)uptake and systemic RNAi mechanisms,and the effective delivery of dsRNA in field conditions.Furthermore,we summarize recent progress in RNAi strategies for crop protection,discuss the advantages and disadvantages of RNAi-based insect control,and propose potential strategies to increase the effectiveness of RNAi in insect control.展开更多
目的建立针对MYOC基因RNAi慢病毒载体的优化及筛选方法,为研究后续突变基因的功能奠定基础。方法 Western Blot筛选RNAi有效靶点,Real-time PCR检测小梁细胞MYOC基因Knock Down效率。结果 Western blotting结果显示含有敲减质粒的组别...目的建立针对MYOC基因RNAi慢病毒载体的优化及筛选方法,为研究后续突变基因的功能奠定基础。方法 Western Blot筛选RNAi有效靶点,Real-time PCR检测小梁细胞MYOC基因Knock Down效率。结果 Western blotting结果显示含有敲减质粒的组别其蛋白表达明显减少;Real-time PCR结果显示在荧光显微镜下观察,与对照组相比含有敲减质粒的组别其MYOC基因的表达明显下降,1#、2#、3#具有确切的干扰效果。结论经过Western blotting及Real-time PCR方法检测所构建的4个干扰靶点的敲减效率,证实RNAi慢病毒干扰效果可靠,且两种方法结果一致,可随机选择1#、2#、3#中的任一位点作为后续干扰靶点。展开更多
基金funded by the National Natural Science Foundation of China(32188102 to Lanqin Xia)Natural Science Foundation for Distinguished Young Scholars of Jiangxi province(20212ACB215001 to Xiudao Yu)+1 种基金supported by the China Scholarship Council(202303250062)the GSCAAS-ULg Joint PhD Program。
文摘RNA interference(RNAi)has been used for agricultural insect pest control based on silencing of targeted insect genes.However,the effectiveness of RNAi and its applications in insect pest control remain challenging.Here we review factors that may affect the effectiveness of RNAi application,including the variability in RNAi efficacy among different insect species,a limited understanding of double-stranded RNA(dsRNA)uptake and systemic RNAi mechanisms,and the effective delivery of dsRNA in field conditions.Furthermore,we summarize recent progress in RNAi strategies for crop protection,discuss the advantages and disadvantages of RNAi-based insect control,and propose potential strategies to increase the effectiveness of RNAi in insect control.
文摘目的建立针对MYOC基因RNAi慢病毒载体的优化及筛选方法,为研究后续突变基因的功能奠定基础。方法 Western Blot筛选RNAi有效靶点,Real-time PCR检测小梁细胞MYOC基因Knock Down效率。结果 Western blotting结果显示含有敲减质粒的组别其蛋白表达明显减少;Real-time PCR结果显示在荧光显微镜下观察,与对照组相比含有敲减质粒的组别其MYOC基因的表达明显下降,1#、2#、3#具有确切的干扰效果。结论经过Western blotting及Real-time PCR方法检测所构建的4个干扰靶点的敲减效率,证实RNAi慢病毒干扰效果可靠,且两种方法结果一致,可随机选择1#、2#、3#中的任一位点作为后续干扰靶点。
