Science:张锋团队今日带来 CRISPR 重磅新应用,核酸检测灵敏度可达单分子

2017-04-14 佚名 学术经纬

今日,知名华人科学家张锋教授与同事在《科学》杂志上发表论文,介绍了一种全新的 CRISPR 应用工具,用以检测核酸。这一工具能快速检测 RNA 或 DNA 分子,灵敏度甚至有望检测出单个核酸。这一重磅研究对于科研与全球公共卫生有着极大的影响。早在 2010 年,科学家们就意识到,CRISPR 平台也许能被用来检测病毒的核酸。但在当时,它的基因编辑潜力过于出众,光芒在一定程度上掩盖了其他领域应用的可

今日,知名华人科学家张锋教授与同事在《科学》杂志上发表论文,介绍了一种全新的 CRISPR 应用工具,用以检测核酸。这一工具能快速检测 RNA 或 DNA 分子,灵敏度甚至有望检测出单个核酸。这一重磅研究对于科研与全球公共卫生有着极大的影响。

早在 2010 年,科学家们就意识到,CRISPR 平台也许能被用来检测病毒的核酸。但在当时,它的基因编辑潜力过于出众,光芒在一定程度上掩盖了其他领域应用的可能性。直到 CRISPR-Cas9 基因编辑技术趋于完善之时,科研人员才有余力检视它在病毒检测上的潜力。

科学家们在今日发表的这个系统与 CRISPR-Cas9 系统有着本质上的差异。它由向导 RNA(gRNA)和 Cas13a 蛋白等部分组成。向导 RNA 能带领 Cas13a 蛋白识别并剪开带有特异序列的 RNA。然而 Cas13a 有一个奇怪的特点,当它切开特异的 RNA 后,就无法管住自己的“双手”,开始疯狂切开它能遇到的任何 RNA。利用这一特性,张锋教授与 James Collins 教授在这套系统里塞入了一种带有 RNA 的荧光标志物。平时它不会发出荧光,但一旦 Cas13a 到处“搞事情”,这种荧光标志物的 RNA 就会被切开,让它发出明亮的光芒。

也就是说,只要设计好合适的向导 RNA,并提供 Cas13a 与荧光标志物,那么一旦这套系统识别到匹配的 RNA 序列,就会发出荧光,告诉研究人员“我们找到了!”

这套系统的灵敏度达到了 50fM(1fM 为 10 的负 15 次方摩尔每升)。

尽管和之前的一些研究相比,它的灵敏度提高了数百万倍,但研究人员依然没有满足。他们想要把灵敏度进一步提高,增加到阿摩尔级(aM,10 的负 18 次方摩尔每升)。为了达到这一目标,他们又用上了“等温扩增”(isothermal amplification)技术,进一步降低了对初始核酸浓度的需求。

在后续的实验中,研究人员发现,使用“等温扩增”技术的这套 CRISPR 系统无论是对 RNA 还是 DNA,灵敏度都达到了单分子级。

这种不遗漏任何线索的特性让研究人员想到了福尔摩斯(Sherlock Holmes)。于是他们杜撰了一个名字,把这套系统称为 SHERLOCK(Specific High Sensitivity Enzymatic Reporter UnLOCKing)。

这套系统开发出来可不是为了好玩。研究人员相信,它能用于各种疾病的检测。首先,研究人员在不同的病毒上测试了它的特异性。实验表明,它能很好地区分两种很类似的寨卡病毒株。这对于准确诊断病情,有效预防疫情爆发而言,有着现实的应用价值。

随后,研究人员又开始将它用于检测癌症突变。在癌症的早期诊断领域,液体活检是一大热门——科学家们希望从血液中分离出专属于癌细胞的突变,从而得知癌症是否开始发作。然而,由于血液中有着大量的非癌细胞 DNA,这一检测过程也是困难无比。在今日发表的论文中,研究团队使用现成的样本,来模拟患者的血液。而实验表明,SHERLOCK 同样能以阿摩尔级的灵敏度,识别出肿瘤特有的突变。

类似的应用还有很多。比如,研究人员用这套系统,成功地鉴定出了人类基因组中的多个单核苷酸多态性(SNP),并能分析出每一个人的特定 SNP 是纯合还是杂合。在这个基因组的时代,SHERLOCK 能得到很广泛的应用。

“能将 Cas13a 应用到灵敏度如此非同寻常的工具中,我感到非常激动。无论对科学还是临床医学,它都是很强大的工具。”张锋教授说。

“我们现在能有效地制作针对任何核酸的探测器,在诊断或研究领域的应用上,这项工具无比强大,”该研究的另一名资深作者 James Collins 教授说道:“这项工具能在患者的血液中寻找到极少量的肿瘤 DNA,帮助研究人员理解肿瘤的突变如何随着时间推移发生改变。另外在公共卫生领域,它也能帮助研究者监控耐药菌的发生频率。科学上它有无限让人激动的可能。”

原始出处:Jonathan S. Gootenberg,Omar O. Abudayyeh1,Jeong Wook Lee,et al.Nucleic acid detection with CRISPR-Cas13a/C2c2.Science  13 Apr 2017:
eaam9321 DOI: 10.1126/science.aam9321

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    2017-12-22 respect
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    2017-05-02 droping

    到底是牛人啊,很强大

    0

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    2017-04-16 yuandd
  5. [GetPortalCommentsPageByObjectIdResponse(id=1775911, encodeId=5d571e7591183, content=<a href='/topic/show?id=ede06694367' target=_blank style='color:#2F92EE;'>#灵敏度#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=28, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=66943, encryptionId=ede06694367, topicName=灵敏度)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=fd6838842412, createdName=respect, createdTime=Fri Dec 22 07:16:00 CST 2017, time=2017-12-22, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1944104, encodeId=01121944104f2, content=<a href='/topic/show?id=be6436145d0' target=_blank style='color:#2F92EE;'>#单分子#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=38, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=36145, encryptionId=be6436145d0, topicName=单分子)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=a12645, createdName=智慧医人, createdTime=Thu Aug 03 13:16:00 CST 2017, time=2017-08-03, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=192982, encodeId=9a0219298208, content=到底是牛人啊,很强大, beContent=null, objectType=article, channel=null, level=null, likeNumber=101, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=1d5e91371, createdName=droping, createdTime=Tue May 02 21:12:46 CST 2017, time=2017-05-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1276018, encodeId=aa7812e601825, content=<a href='/topic/show?id=755d5211a0' target=_blank style='color:#2F92EE;'>#CRISPR#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=31, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=5211, encryptionId=755d5211a0, topicName=CRISPR)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=583c133, createdName=yuandd, createdTime=Sun Apr 16 11:16:00 CST 2017, time=2017-04-16, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1375552, encodeId=189513e5552b6, content=<a href='/topic/show?id=2949159989d' target=_blank style='color:#2F92EE;'>#SCIE#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=38, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=15998, encryptionId=2949159989d, topicName=SCIE)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=17a3387, createdName=jichang, createdTime=Sun Apr 16 11:16:00 CST 2017, time=2017-04-16, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1549097, encodeId=f641154909e03, content=<a href='/topic/show?id=f7879600061' target=_blank style='color:#2F92EE;'>#重磅#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=96000, encryptionId=f7879600061, topicName=重磅)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=3acc14121561, createdName=xxxx1054, createdTime=Sun Apr 16 11:16:00 CST 2017, time=2017-04-16, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=187474, encodeId=c3ea18e47474, content=涨知识,学习了, beContent=null, objectType=article, channel=null, level=null, likeNumber=132, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://img.medsci.cn/20220115/46bcf39c32de4aa6b45c5f9d66c8ee77/6cb4a20c55bb4b7691122f47747bfca2.jpg, createdBy=9dad1662329, createdName=1ddf0692m34(暂无匿称), createdTime=Fri Apr 14 22:50:58 CST 2017, time=2017-04-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=187467, encodeId=1be218e46714, content=这项工具能在患者的血液中寻找到极少量的肿瘤 DNA,帮助研究人员理解肿瘤的突变如何随着时间推移发生改变, beContent=null, objectType=article, channel=null, level=null, likeNumber=97, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=fd301464769, createdName=Albert Wu, createdTime=Fri Apr 14 21:59:12 CST 2017, time=2017-04-14, status=1, ipAttribution=)]
    2017-04-16 jichang
  6. [GetPortalCommentsPageByObjectIdResponse(id=1775911, encodeId=5d571e7591183, content=<a href='/topic/show?id=ede06694367' target=_blank style='color:#2F92EE;'>#灵敏度#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=28, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=66943, encryptionId=ede06694367, topicName=灵敏度)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=fd6838842412, createdName=respect, createdTime=Fri Dec 22 07:16:00 CST 2017, time=2017-12-22, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1944104, encodeId=01121944104f2, content=<a href='/topic/show?id=be6436145d0' target=_blank style='color:#2F92EE;'>#单分子#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=38, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=36145, encryptionId=be6436145d0, topicName=单分子)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=a12645, createdName=智慧医人, createdTime=Thu Aug 03 13:16:00 CST 2017, time=2017-08-03, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=192982, encodeId=9a0219298208, content=到底是牛人啊,很强大, beContent=null, objectType=article, channel=null, level=null, likeNumber=101, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=1d5e91371, createdName=droping, createdTime=Tue May 02 21:12:46 CST 2017, time=2017-05-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1276018, encodeId=aa7812e601825, content=<a href='/topic/show?id=755d5211a0' target=_blank style='color:#2F92EE;'>#CRISPR#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=31, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=5211, encryptionId=755d5211a0, topicName=CRISPR)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=583c133, createdName=yuandd, createdTime=Sun Apr 16 11:16:00 CST 2017, time=2017-04-16, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1375552, encodeId=189513e5552b6, content=<a href='/topic/show?id=2949159989d' target=_blank style='color:#2F92EE;'>#SCIE#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=38, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=15998, encryptionId=2949159989d, topicName=SCIE)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=17a3387, createdName=jichang, createdTime=Sun Apr 16 11:16:00 CST 2017, time=2017-04-16, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1549097, encodeId=f641154909e03, content=<a href='/topic/show?id=f7879600061' target=_blank style='color:#2F92EE;'>#重磅#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=96000, encryptionId=f7879600061, topicName=重磅)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=3acc14121561, createdName=xxxx1054, createdTime=Sun Apr 16 11:16:00 CST 2017, time=2017-04-16, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=187474, encodeId=c3ea18e47474, content=涨知识,学习了, beContent=null, objectType=article, channel=null, level=null, likeNumber=132, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://img.medsci.cn/20220115/46bcf39c32de4aa6b45c5f9d66c8ee77/6cb4a20c55bb4b7691122f47747bfca2.jpg, createdBy=9dad1662329, createdName=1ddf0692m34(暂无匿称), createdTime=Fri Apr 14 22:50:58 CST 2017, time=2017-04-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=187467, encodeId=1be218e46714, content=这项工具能在患者的血液中寻找到极少量的肿瘤 DNA,帮助研究人员理解肿瘤的突变如何随着时间推移发生改变, beContent=null, objectType=article, channel=null, level=null, likeNumber=97, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=fd301464769, createdName=Albert Wu, createdTime=Fri Apr 14 21:59:12 CST 2017, time=2017-04-14, status=1, ipAttribution=)]
    2017-04-16 xxxx1054
  7. [GetPortalCommentsPageByObjectIdResponse(id=1775911, encodeId=5d571e7591183, content=<a href='/topic/show?id=ede06694367' target=_blank style='color:#2F92EE;'>#灵敏度#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=28, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=66943, encryptionId=ede06694367, topicName=灵敏度)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=fd6838842412, createdName=respect, createdTime=Fri Dec 22 07:16:00 CST 2017, time=2017-12-22, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1944104, encodeId=01121944104f2, content=<a href='/topic/show?id=be6436145d0' target=_blank style='color:#2F92EE;'>#单分子#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=38, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=36145, encryptionId=be6436145d0, topicName=单分子)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=a12645, createdName=智慧医人, createdTime=Thu Aug 03 13:16:00 CST 2017, time=2017-08-03, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=192982, encodeId=9a0219298208, content=到底是牛人啊,很强大, beContent=null, objectType=article, channel=null, level=null, likeNumber=101, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=1d5e91371, createdName=droping, createdTime=Tue May 02 21:12:46 CST 2017, time=2017-05-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1276018, encodeId=aa7812e601825, content=<a href='/topic/show?id=755d5211a0' target=_blank style='color:#2F92EE;'>#CRISPR#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=31, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=5211, encryptionId=755d5211a0, topicName=CRISPR)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=583c133, createdName=yuandd, createdTime=Sun Apr 16 11:16:00 CST 2017, time=2017-04-16, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1375552, encodeId=189513e5552b6, content=<a href='/topic/show?id=2949159989d' target=_blank style='color:#2F92EE;'>#SCIE#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=38, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=15998, encryptionId=2949159989d, topicName=SCIE)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=17a3387, createdName=jichang, createdTime=Sun Apr 16 11:16:00 CST 2017, time=2017-04-16, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1549097, encodeId=f641154909e03, content=<a href='/topic/show?id=f7879600061' target=_blank style='color:#2F92EE;'>#重磅#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=96000, encryptionId=f7879600061, topicName=重磅)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=3acc14121561, createdName=xxxx1054, createdTime=Sun Apr 16 11:16:00 CST 2017, time=2017-04-16, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=187474, encodeId=c3ea18e47474, content=涨知识,学习了, beContent=null, objectType=article, channel=null, level=null, likeNumber=132, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://img.medsci.cn/20220115/46bcf39c32de4aa6b45c5f9d66c8ee77/6cb4a20c55bb4b7691122f47747bfca2.jpg, createdBy=9dad1662329, createdName=1ddf0692m34(暂无匿称), createdTime=Fri Apr 14 22:50:58 CST 2017, time=2017-04-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=187467, encodeId=1be218e46714, content=这项工具能在患者的血液中寻找到极少量的肿瘤 DNA,帮助研究人员理解肿瘤的突变如何随着时间推移发生改变, beContent=null, objectType=article, channel=null, level=null, likeNumber=97, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=fd301464769, createdName=Albert Wu, createdTime=Fri Apr 14 21:59:12 CST 2017, time=2017-04-14, status=1, ipAttribution=)]
    2017-04-14 1ddf0692m34(暂无匿称)

    涨知识,学习了

    0

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    2017-04-14 Albert Wu

    这项工具能在患者的血液中寻找到极少量的肿瘤 DNA,帮助研究人员理解肿瘤的突变如何随着时间推移发生改变

    0

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