目的结合TargetScan软件预测,体外验证小鼠microRNA-135a(miR-135a)与信号传导及转录激活因子6(signal transducer and activator of transcription 6,STAT6)的靶点结合作用。方法将聚合酶链反应(polymerase chain reaction,PCR)扩增的...目的结合TargetScan软件预测,体外验证小鼠microRNA-135a(miR-135a)与信号传导及转录激活因子6(signal transducer and activator of transcription 6,STAT6)的靶点结合作用。方法将聚合酶链反应(polymerase chain reaction,PCR)扩增的靶基因STAT6的非编码3'UTR和突变基因分别克隆到双荧光素酶报告载体中,构建野生型和突变型载体。分别将miR-135a模拟物、模拟对照物与靶基因野生型、突变型质粒载体共转染体外培养的293T细胞,测定载体的校正荧光hluc值及报告荧光hRluc值。结果与模拟对照物相比较,miR-135a模拟物与靶基因STAT6的3'UTR野生型质粒的相互作用能明显降低hRluc/hluc的相对荧光比值,差异有统计学意义(P<0.01);而miR-135a模拟物与突变型质粒相作用则对hRluc/hluc的相对荧光比值比较差异无统计学意义(P>0.05)。结论小鼠miR-135a与STAT6的3'UTR体外特异性结合作用,为进一步探讨小鼠miR-135a对变应性鼻炎的免疫调控机制提供了依据。展开更多
Long noncoding RNAs （IncRNAs） participate in the regulation of multiple biological processes via diverse manners, one of which is functioning as endogenous target mimics （eTMs） to modulate microRNAs （miRNAs） by ...Long noncoding RNAs （IncRNAs） participate in the regulation of multiple biological processes via diverse manners, one of which is functioning as endogenous target mimics （eTMs） to modulate microRNAs （miRNAs） by competing for their targets. Previously, we have predicted one IncRNA （osa-eTM160） as an endogenous repressor of osa-miR160 and validated the target mimicry ability of osa-eTM160 for ath-miR160 in Arabidopsis thaliana, yet the functions of osa-eTM160 in rice remain obscure. Here, we demonstrated that osa-eTM160 attenuated the repression of osa-miR160 on osa-ARF18 mRNAs during early anther developmental stages through the target mimicry manner, therefore to regulate rice seed setting and seed size. These findings revealed the roles of osa-eTM160 in rice, and indicated that lncRNAs with eTM functions may serve as temporal regulators to modulate the effects of miRNAs at specific developmental stages.展开更多
文摘目的结合TargetScan软件预测,体外验证小鼠microRNA-135a(miR-135a)与信号传导及转录激活因子6(signal transducer and activator of transcription 6,STAT6)的靶点结合作用。方法将聚合酶链反应(polymerase chain reaction,PCR)扩增的靶基因STAT6的非编码3'UTR和突变基因分别克隆到双荧光素酶报告载体中,构建野生型和突变型载体。分别将miR-135a模拟物、模拟对照物与靶基因野生型、突变型质粒载体共转染体外培养的293T细胞,测定载体的校正荧光hluc值及报告荧光hRluc值。结果与模拟对照物相比较,miR-135a模拟物与靶基因STAT6的3'UTR野生型质粒的相互作用能明显降低hRluc/hluc的相对荧光比值,差异有统计学意义(P<0.01);而miR-135a模拟物与突变型质粒相作用则对hRluc/hluc的相对荧光比值比较差异无统计学意义(P>0.05)。结论小鼠miR-135a与STAT6的3'UTR体外特异性结合作用,为进一步探讨小鼠miR-135a对变应性鼻炎的免疫调控机制提供了依据。
基金supported by the National Natural Science Foundation of China(31371318 to M.W.)the State Key Laboratory of Plant Genomics(SKLPG2011B0105 to X.-J.W.)Youth Innovation Promotion Association Grant from Chinese Academy of Sciences(to M.W.)
文摘Long noncoding RNAs （IncRNAs） participate in the regulation of multiple biological processes via diverse manners, one of which is functioning as endogenous target mimics （eTMs） to modulate microRNAs （miRNAs） by competing for their targets. Previously, we have predicted one IncRNA （osa-eTM160） as an endogenous repressor of osa-miR160 and validated the target mimicry ability of osa-eTM160 for ath-miR160 in Arabidopsis thaliana, yet the functions of osa-eTM160 in rice remain obscure. Here, we demonstrated that osa-eTM160 attenuated the repression of osa-miR160 on osa-ARF18 mRNAs during early anther developmental stages through the target mimicry manner, therefore to regulate rice seed setting and seed size. These findings revealed the roles of osa-eTM160 in rice, and indicated that lncRNAs with eTM functions may serve as temporal regulators to modulate the effects of miRNAs at specific developmental stages.
