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CRISPR、RNAi、TALEN一张图教你做出正确选择

2015-05-26 叶予 生物通

研究基因功能最常见的方法是,减少或者阻断基因表达,然后进行表型分析。十多年来,RNAi一直是这一领域的王者,然而新兴技术的涌现(尤其是CRISPR技术)正在逐渐瓦解RNAi的统治地位。日新月异的技术发展为生物学研究提供了越来越大的助力,也给研究者们带来了一个有些纠结的问题,“到底应该选择那一种技术呢”。  1998年Andrew Fire 和Craig Mello两位科学家首次

研究基因功能最常见的方法是,减少或者阻断基因表达,然后进行表型分析。十多年来,RNAi一直是这一领域的王者,然而新兴技术的涌现(尤其是CRISPR技术)正在逐渐瓦解RNAi的统治地位。日新月异的技术发展为生物学研究提供了越来越大的助力,也给研究者们带来了一个有些纠结的问题,“到底应该选择那一种技术呢”。 

1998年Andrew Fire 和Craig Mello两位科学家首次在线虫中证明了RNAi的存在,他们也因此获得了诺贝尔生理/医学奖。2001年马普研究所的科学家们又在哺乳动物中发现了RNAi特异性沉默机制。这一技术随即成为了反向遗传学的重要工具,被视为靶向任何基因的“神奇子弹”。虽然RNAi是一种普遍采用的简单技术,但它属于基因下调(Knockdown)方法,不能完全去除基因的功能。

真正帮助人们实现完全功能缺失的是锌指酶ZFN和TALEN(transcription activator-like effector nucleases)。这些技术使用了可以识别特定DNA序列的DNA结合结构域(DBD),融合了核酸酶的DBD可以向目的基因引入双链断裂(DSB)和突变,最终敲除基因。不过,ZFN和TALEN的光芒很快就被后起之秀CRISPR/Cas所掩盖。 

CRISPR/Cas原本是细菌抵御病毒的重要武器,现在它已经成为了基因组编辑的强大工具。风头正劲的CRISPR/Cas不仅操作简便,而且还有着很强的可扩展性,被广泛应用到各种生物中,催生了大量的研究成果。根据功能元件的不同,CRISPR/Cas系统可以分为I类系统、II类系统和III类系统。目前使用最为广泛的是酿脓链球菌的II型CRISPR/Cas9,它已经被成功用于编辑人类基因组。

最新一期Molecular Cell杂志推出了技术特刊,介绍了生物学领域近年来出现的新兴技术。其中一篇文章对RNAi、TALEN和CRISPR这三大工具的核心技术进行了全面比较,并且为基因功能研究提供了一份实用指南。(现在这篇文章可以免费下载)研究者们可以根据自己的需要,简单直观的找到最适合自己的技术。 

RNAi、TALEN和CRISPR选择指南(见下图)


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    2016-02-04 yinxm8315
  2. [GetPortalCommentsPageByObjectIdResponse(id=1751653, encodeId=446f1e5165322, content=<a href='/topic/show?id=2c2c1553e56' target=_blank style='color:#2F92EE;'>#RNAi#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=47, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=15537, encryptionId=2c2c1553e56, topicName=RNAi)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=d9d436366397, createdName=yinxm8315, createdTime=Thu Feb 04 04:21:00 CST 2016, time=2016-02-04, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=25635, encodeId=76ce2563504, content=赞, beContent=null, objectType=article, channel=null, level=null, likeNumber=131, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=ee0d1618732, createdName=huaxipanxing, createdTime=Sun May 31 21:36:00 CST 2015, time=2015-05-31, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1275833, encodeId=d98112e5833fc, content=<a href='/topic/show?id=755d5211a0' target=_blank style='color:#2F92EE;'>#CRISPR#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=44, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=5211, encryptionId=755d5211a0, topicName=CRISPR)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=583c133, createdName=yuandd, createdTime=Thu May 28 07:21:00 CST 2015, time=2015-05-28, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=24973, encodeId=e823249e392, content=真正的学习了!, beContent=null, objectType=article, channel=null, level=null, likeNumber=177, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://api.medsci.cn/resource/upload/20150526/IMG5564536F3C41D5237.jpg, createdBy=4c0c1612254, createdName=summer474cn, createdTime=Tue May 26 19:41:00 CST 2015, time=2015-05-26, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=24958, encodeId=4ab72495846, content=赞!, beContent=null, objectType=article, channel=null, level=null, likeNumber=129, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://thirdwx.qlogo.cn/mmopen/vi_32/D26s4tMwZbRicDx7kCqw3ZhsUu5Xqw61EKDTCNyMwj0DnclBCmVmD1q3xJ8dkLEibbgBgkGicFEEB6kviags8Dar4A/132, createdBy=857c100601, createdName=lisalin, createdTime=Tue May 26 16:52:00 CST 2015, time=2015-05-26, status=1, ipAttribution=)]
    2015-05-31 huaxipanxing

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  3. [GetPortalCommentsPageByObjectIdResponse(id=1751653, encodeId=446f1e5165322, content=<a href='/topic/show?id=2c2c1553e56' target=_blank style='color:#2F92EE;'>#RNAi#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=47, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=15537, encryptionId=2c2c1553e56, topicName=RNAi)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=d9d436366397, createdName=yinxm8315, createdTime=Thu Feb 04 04:21:00 CST 2016, time=2016-02-04, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=25635, encodeId=76ce2563504, content=赞, beContent=null, objectType=article, channel=null, level=null, likeNumber=131, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=ee0d1618732, createdName=huaxipanxing, createdTime=Sun May 31 21:36:00 CST 2015, time=2015-05-31, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1275833, encodeId=d98112e5833fc, content=<a href='/topic/show?id=755d5211a0' target=_blank style='color:#2F92EE;'>#CRISPR#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=44, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=5211, encryptionId=755d5211a0, topicName=CRISPR)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=583c133, createdName=yuandd, createdTime=Thu May 28 07:21:00 CST 2015, time=2015-05-28, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=24973, encodeId=e823249e392, content=真正的学习了!, beContent=null, objectType=article, channel=null, level=null, likeNumber=177, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://api.medsci.cn/resource/upload/20150526/IMG5564536F3C41D5237.jpg, createdBy=4c0c1612254, createdName=summer474cn, createdTime=Tue May 26 19:41:00 CST 2015, time=2015-05-26, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=24958, encodeId=4ab72495846, content=赞!, beContent=null, objectType=article, channel=null, level=null, likeNumber=129, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://thirdwx.qlogo.cn/mmopen/vi_32/D26s4tMwZbRicDx7kCqw3ZhsUu5Xqw61EKDTCNyMwj0DnclBCmVmD1q3xJ8dkLEibbgBgkGicFEEB6kviags8Dar4A/132, createdBy=857c100601, createdName=lisalin, createdTime=Tue May 26 16:52:00 CST 2015, time=2015-05-26, status=1, ipAttribution=)]
    2015-05-28 yuandd
  4. [GetPortalCommentsPageByObjectIdResponse(id=1751653, encodeId=446f1e5165322, content=<a href='/topic/show?id=2c2c1553e56' target=_blank style='color:#2F92EE;'>#RNAi#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=47, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=15537, encryptionId=2c2c1553e56, topicName=RNAi)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=d9d436366397, createdName=yinxm8315, createdTime=Thu Feb 04 04:21:00 CST 2016, time=2016-02-04, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=25635, encodeId=76ce2563504, content=赞, beContent=null, objectType=article, channel=null, level=null, likeNumber=131, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=ee0d1618732, createdName=huaxipanxing, createdTime=Sun May 31 21:36:00 CST 2015, time=2015-05-31, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1275833, encodeId=d98112e5833fc, content=<a href='/topic/show?id=755d5211a0' target=_blank style='color:#2F92EE;'>#CRISPR#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=44, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=5211, encryptionId=755d5211a0, topicName=CRISPR)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=583c133, createdName=yuandd, createdTime=Thu May 28 07:21:00 CST 2015, time=2015-05-28, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=24973, encodeId=e823249e392, content=真正的学习了!, beContent=null, objectType=article, channel=null, level=null, likeNumber=177, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://api.medsci.cn/resource/upload/20150526/IMG5564536F3C41D5237.jpg, createdBy=4c0c1612254, createdName=summer474cn, createdTime=Tue May 26 19:41:00 CST 2015, time=2015-05-26, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=24958, encodeId=4ab72495846, content=赞!, beContent=null, objectType=article, channel=null, level=null, likeNumber=129, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://thirdwx.qlogo.cn/mmopen/vi_32/D26s4tMwZbRicDx7kCqw3ZhsUu5Xqw61EKDTCNyMwj0DnclBCmVmD1q3xJ8dkLEibbgBgkGicFEEB6kviags8Dar4A/132, createdBy=857c100601, createdName=lisalin, createdTime=Tue May 26 16:52:00 CST 2015, time=2015-05-26, status=1, ipAttribution=)]
    2015-05-26 summer474cn

    真正的学习了!

    0

  5. [GetPortalCommentsPageByObjectIdResponse(id=1751653, encodeId=446f1e5165322, content=<a href='/topic/show?id=2c2c1553e56' target=_blank style='color:#2F92EE;'>#RNAi#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=47, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=15537, encryptionId=2c2c1553e56, topicName=RNAi)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=d9d436366397, createdName=yinxm8315, createdTime=Thu Feb 04 04:21:00 CST 2016, time=2016-02-04, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=25635, encodeId=76ce2563504, content=赞, beContent=null, objectType=article, channel=null, level=null, likeNumber=131, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=ee0d1618732, createdName=huaxipanxing, createdTime=Sun May 31 21:36:00 CST 2015, time=2015-05-31, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1275833, encodeId=d98112e5833fc, content=<a href='/topic/show?id=755d5211a0' target=_blank style='color:#2F92EE;'>#CRISPR#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=44, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=5211, encryptionId=755d5211a0, topicName=CRISPR)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=583c133, createdName=yuandd, createdTime=Thu May 28 07:21:00 CST 2015, time=2015-05-28, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=24973, encodeId=e823249e392, content=真正的学习了!, beContent=null, objectType=article, channel=null, level=null, likeNumber=177, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://api.medsci.cn/resource/upload/20150526/IMG5564536F3C41D5237.jpg, createdBy=4c0c1612254, createdName=summer474cn, createdTime=Tue May 26 19:41:00 CST 2015, time=2015-05-26, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=24958, encodeId=4ab72495846, content=赞!, beContent=null, objectType=article, channel=null, level=null, likeNumber=129, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://thirdwx.qlogo.cn/mmopen/vi_32/D26s4tMwZbRicDx7kCqw3ZhsUu5Xqw61EKDTCNyMwj0DnclBCmVmD1q3xJ8dkLEibbgBgkGicFEEB6kviags8Dar4A/132, createdBy=857c100601, createdName=lisalin, createdTime=Tue May 26 16:52:00 CST 2015, time=2015-05-26, status=1, ipAttribution=)]
    2015-05-26 lisalin

    赞!

    0

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