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Nature Methods:便携、低成本的单细胞RNA测序新手段

2017-02-20 佚名 生物探索

每个细胞的基因表达谱随其功能而异。从血液或组织样本中的许多单个细胞中提取RNA可以根据基因表达模式区分细胞。这使科学家有机会确定细胞功能,包括它们在疾病中的作用或对治疗的反应。MIT的研究人员已经开发出一种便携式技术,可以快速制备多种细胞的RNA同时进行测序,他们相信这种方法将会更广泛地被应用。新的技术被称为Seq-Well,可以让科学家在组织样本中更容易识别不同类型的细胞,帮助他们研究免疫细胞如



每个细胞的基因表达谱随其功能而异。从血液或组织样本中的许多单个细胞中提取RNA可以根据基因表达模式区分细胞。这使科学家有机会确定细胞功能,包括它们在疾病中的作用或对治疗的反应。

MIT的研究人员已经开发出一种便携式技术,可以快速制备多种细胞的RNA同时进行测序,他们相信这种方法将会更广泛地被应用。新的技术被称为Seq-Well,可以让科学家在组织样本中更容易识别不同类型的细胞,帮助他们研究免疫细胞如何对抗感染和癌细胞对治疗的反应以及其它的应用。

1单细胞测序的难点

对大量细胞进行单细胞测序的关键是保持跟踪哪些RNA转录本来自哪个细胞。这需要将细胞分开在单个管或多孔板格中,然后分别转化成测序文库。

这个过程用利用毛细管直接吸取细胞内容物的显微操作可以进行,但不能处理含有数千个细胞大样本,如血液样本或组织活检,且成本在25元至35美元/每细胞。Shalek等人最近开发的微流控技术来让分选过程自动化和平行化,尽管微流控技术最近在生命科学领域有革命性作用,但所需的设备量使得它不可能被轻易搬运。

2作者对发明的介绍

通讯作者Alex K. Shalek说:“不是选择用一个标记定义的细胞类型,而用单细胞的RNA测序,我们可以看看某个细胞在特定时刻的表达。”

Shalek和他的同事们在过去几年的发展单细胞RNA测序策略。在这项新的研究中,他与MIT的化学工程学副教授J. Christopher Love合作创建了一种新的技术,可以使用非常简单的设备快速分析大量的细胞。

Love博士说:“我们将Shalek的技术和我们的结合起来,是研究人员能够在不同的临床样本和设备的范畴中进行这种测序。这种方法克服了现在的一些障碍,能让大家更广泛地采用这些技术。这是一个简单的系统并带到能到世界各地去,我认为它有着难以置信的力量。”

3该技术具有快速分析能力

在过去的几年里,Love的实验室开发出一种微型系统,能分离出单个的细胞和检测每个细胞分泌的抗体和其它蛋白。这个设备类似于一个小型的冰托盘,每个细胞有独立的隔间。Love开发了一个叫做microengraving的过程使用这些托盘。这些托盘可以容纳数以万计的细胞,来检测每个细胞蛋白质的分泌。



Shalek 和Love这两个在一起合作的研究者,意识到Love开发的从单细胞分析其蛋白分泌的技术能够用于设计一个便携式设备快速、廉价地来进行单细胞RNA测序。

使用此方法对RNA进行测序,研究人员建立了捕捉单个细胞的纳米隔间阵列,用有条码的磁珠捕获RNA片段。每个纳米隔间用半透膜封闭允许化学信号通过。当RNA结合到磁珠上后,将其移除并测序。使用这个过程,每个单元格的成本小于1美元。

4
用此技术揭开未知

类似以前的单细胞RNA测序技术,Seq-well过程能够很好捕获和分析序列约每个细胞10至15%的RNA转录。

在这篇文章中,研究者使用Seq-Well对被称为巨噬细胞的免疫细胞进行分析,这些巨噬细胞来源于感染了肺结核的样本。这个新方法让他们能够识别不同的现有巨噬细胞群体的对感染的反应。Shalek和他的实验室成员也将这个技术带到了南非,在那里分析结核病人和HIV感染患者的样本。

Love的实验室现在使用这种方法来分析人的免疫细胞与食物过敏,这可能有助于研究人员确定为什么有些人更可能对治疗他们过敏的疗法反应良好。Love认为在慢性病中仍有许多未知因素,这些类型的工具帮助研究人员发现新的见解。

研究小组还联手与Dana Farber /哈佛癌症中心的临床研究者,将此技术应用于新的组合疗法治疗癌症。

原始出处:

Todd M Gierahn, Marc H Wadsworth II,et all.Seq-Well: portable, low-cost RNA sequencing of single cells at high throughput.Nature Methods (2017) doi:10.1038/nmeth.4179

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    2017-05-31 liye789132251
  2. [GetPortalCommentsPageByObjectIdResponse(id=1881674, encodeId=222218816e443, content=<a href='/topic/show?id=3b2112532d8' target=_blank style='color:#2F92EE;'>#Nat#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=54, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=12532, encryptionId=3b2112532d8, topicName=Nat)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=2e6f107, createdName=liye789132251, createdTime=Wed May 31 15:33:00 CST 2017, time=2017-05-31, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1895994, encodeId=1221189599457, content=<a href='/topic/show?id=4a391158248' target=_blank style='color:#2F92EE;'>#MET#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=0, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=11582, encryptionId=4a391158248, topicName=MET)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=18fc139, createdName=一闲, createdTime=Tue Sep 12 21:33:00 CST 2017, time=2017-09-12, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=177571, encodeId=43a21e757124, content=很好的研究。, beContent=null, objectType=article, channel=null, level=null, likeNumber=75, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/NUyjXTCJjo7LIiaRZTzJ3SEfiauCzTMPW7YvPPYLNJBXG9oh6Al1icq2VQQkIHWxqXehcicTS62YKJVBhxeth3wggw/0, createdBy=7ce01621306, createdName=天涯183, createdTime=Thu Feb 23 09:19:57 CST 2017, time=2017-02-23, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1418917, encodeId=090a141891e3f, content=<a href='/topic/show?id=abde36296f5' target=_blank style='color:#2F92EE;'>#单细胞RNA测序#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=47, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=36296, encryptionId=abde36296f5, topicName=单细胞RNA测序)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=eda53444564, createdName=wetgdt, createdTime=Wed Feb 22 10:33:00 CST 2017, time=2017-02-22, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1516807, encodeId=e9ba151680e4c, content=<a href='/topic/show?id=eb451556289' target=_blank style='color:#2F92EE;'>#RNA测序#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=62, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=15562, encryptionId=eb451556289, topicName=RNA测序)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=3b3411016716, createdName=ms24272190615788285182, createdTime=Wed Feb 22 10:33:00 CST 2017, time=2017-02-22, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=177241, encodeId=28f61e72417a, content=这个发明前途无量, beContent=null, objectType=article, channel=null, level=null, likeNumber=86, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://cdnapi.center.medsci.cn/medsci/head/2017/04/24/1c2dbd6706e64df33befd6663f1d6b37.jpg, createdBy=94f61661779, createdName=lyh994, createdTime=Tue Feb 21 09:09:17 CST 2017, time=2017-02-21, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=177236, encodeId=b2ae1e7236b2, content=非常好的技术, beContent=null, objectType=article, channel=null, level=null, likeNumber=64, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/NUyjXTCJjo7LIiaRZTzJ3SEfiauCzTMPW7YvPPYLNJBXG9oh6Al1icq2VQQkIHWxqXehcicTS62YKJVBhxeth3wggw/0, createdBy=7ce01621306, createdName=天涯183, createdTime=Tue Feb 21 08:55:49 CST 2017, time=2017-02-21, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=177200, encodeId=fdee1e7200ad, content=好好努力, beContent=null, objectType=article, channel=null, level=null, likeNumber=72, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=45101626056, createdName=忠诚向上, createdTime=Tue Feb 21 06:45:08 CST 2017, time=2017-02-21, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=177174, encodeId=fba41e71745d, content=学习了赞一个, beContent=null, objectType=article, channel=null, level=null, likeNumber=81, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://cacheapi.medsci.cn/resource/upload/20160905/IMG57CD39B82D12E7926.jpg, createdBy=ac2c1946830, createdName=青龙偃月, createdTime=Mon Feb 20 22:45:20 CST 2017, time=2017-02-20, status=1, ipAttribution=)]
    2017-09-12 一闲
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    2017-02-23 天涯183

    很好的研究。

    0

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    2017-02-21 lyh994

    这个发明前途无量

    0

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    2017-02-21 天涯183

    非常好的技术

    0

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    2017-02-21 忠诚向上

    好好努力

    0

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    2017-02-20 青龙偃月

    学习了赞一个

    0

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