? 浣曞窛/璐炬鑺宠棰樼粍鍦ˋngew. Chem. Int. Ed. 鎶ラ亾RNA淇グm6A鍗曞熀鍥犲崟纰卞熀妫€娴嬫柊鎶€鏈?--鐢熺墿鏈夋満涓庡垎瀛愬伐绋嬫暀鑲查儴閲嶇偣瀹為獙瀹?

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何川/贾桂芳课题组在Angew. Chem. Int. Ed. 报道RNA修饰m6A单基因单碱基检测新技?/h1>
作者:     来源?nbsp;    发布日期:2018-10-25

2018?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">10月,北京大学化学与分子工程学院何?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">/贾桂芳课题组在?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">Angew. Chem. Int. Ed. (德国应用化学)》杂志在线发表题为?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">An Elongation and Ligation-Based qPCR Amplification Method for the Radiolabeling-Free Detection of Locus-Specific N6-methyladenosine Modifications?/span>的研究论?span> (DOI:10.1002/anie.201807942)。文章报道了一种快捷检?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">RNA?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">N6-甲基腺嘌呤(N6-methyladenosine?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">m6A)的新方法?/p>

表观转录组学?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">epitranscriptomics,又称?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">RNA表观遗传学”)是近年来兴起的前沿科研领域?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">RNA化学修饰m6A作为表观转录组学中的明星修饰,是真核mRNA和非编码RNA中的主要化学修饰?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">m6A可以被修饰酶和去修饰酶进行动态可逆调节,并被结合蛋白识别调控RNA加工代谢,参与许多重要生物学调节,如周期节律、干细胞分化、癌症发生发展及RNA病毒等?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">m6A检测对于其分子生物学功能和相关疾病研究非常重要。目前报道的检测方法中只有SCARLET方法能够实现?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">mRNA?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">lncRNA?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">m6A单碱基分辨率的检测,但该方法操作复杂耗时,需要较大剂量的放射性同位素标记,难以广泛应用?/p>

北京大学化学学院何川/贾桂芳课题组开发了SELECT方法,原理如?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">1所示,该方法简单快捷,用时只需三小时,能够实现低丰度转录本?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">m6A单碱基分辨率的检测。该方法基于m6A修饰的两点特性:?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">i)能够阻?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">DNA聚合酶在反转录过程的单碱基延伸;?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">ii)能够降低缺口连接酶的连接效率?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">SELECT方法采用荧光定量PCR的方法进行定量。结合方法优化,发展?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">SELECT方法?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">3个应用实例:?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">i)在生物学样本?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">RNA或?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">mRNA中检?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">mRNA?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">lncRNA上单位点是否含有m6A修饰;(ii)检测生物学样本中特?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">m6A位点的修饰比例;?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">iii)鉴?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">m6A甲基转移酶或去甲基酶的生理作用靶点?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">SELECT方法不仅简化了生物学样品中m6A修饰的检测过程,实现低丰?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">RNA?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">m6A单碱基分辨率的检测,还能够应用于其他RNA化学修饰的检测中,例?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">N1-甲基腺嘌呤(N1-methyladenosine?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">m1A)和2?O-甲基化修饰(2?O-methylation?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">Nm)等?/p>

?/span>1. SELECT方法原理示意?/span>

北京大学化学与分子工程学?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">14级博士研究生肖雨同学为第一作者,贾桂芳副研究员为本文通讯作者,相关工作得到了国家科技部和国家自然科学基金委等经费资助?/p>

原文链接?span style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; font-family: Calibri, serif; padding-top: 0px; -webkit-font-smoothing: subpixel-antialiased">//doi.org/10.1002/anie.201807942

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