Nature: 无所不能的CRISPR,居然还可用于疾病检测

2017-04-16 MedSci MedSci原创

细菌适应性免疫系统利用CRISPRs(短回文重复序列间隔聚集)和CRISPR相关(CAS)的蛋白质行使RNA指导的核酸切割。虽然大多数原核生物适应性免疫系统通常以DNA为靶底物,但是III型和VI CRISPR系统直接对单链RNA底物干扰。在VI的系统中,单亚基C2C2蛋白用作为RNA引导的RNA内切酶(RNA酶)。这种酶如何获得免疫监测所必需的成熟CRISPR RNA(crRNA),以及如何进行

CRISPR因能高效地敲除及插入基因,被誉为“基因魔剪”,风光无限,然而,近日,来自美国加州大学伯克利分校的研究人员报道了CRISPR在疾病诊断中的应用,相关研究成果刊登于国际杂志Nature上。

细菌适应性免疫系统利用CRISPRs(短回文重复序列间隔聚集)和CRISPR相关(CAS)的蛋白质行使RNA指导的核酸切割。虽然大多数原核生物适应性免疫系统通常以DNA为靶底物,但是III型和VI CRISPR系统直接对单链RNA底物干扰。在VI的系统中,单亚基C2C2蛋白用作为RNA引导的RNA内切酶(RNA酶)。这种酶如何获得免疫监测所必需的成熟CRISPR RNAcrRNA),以及如何进行crRNA介导的RNA切割仍不清楚。比如,寨卡病毒和登革热病毒因临床症状相似而很难辨别,而在SHERLOCK的帮助下,医生可以很快确认患者到底是感染了哪种病毒。此外,就算两个病毒之间只存在单个核苷酸上的差别,SHERLOCK也能将其识别出来,这就可以区别来自非洲和来自美洲的寨卡病毒。

这里研究人员通过研究显示细菌C2c2与其RNA激活的单链RNA降解活性不同,负责CRISPR RNA成熟时具有独特的核糖核酸酶活性。这些双重RNA酶功能在化学和机械上彼此不同,并且与进化上无关的CRISPRCpf1crRNA加工行为也不同。C2c2的两个RNA酶活性使得能够同时进行多个处理和加载引导RNA,从而允许对细胞转录本进行敏感的检测。

原始出处:

Alexandra East-Seletsky, Mitchell R. O’Connell, et.al. Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection.Nature 538, 270–273 (13 October 2016) doi:10.1038/nature19802

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    2017-04-18 yuandd