Commun Bio:用共价键提高CRISPR/Cas9的修复效率

2018-08-25 Nature自然科研 Nature自然科研

近日在《通讯-生物学》发表的一项名为Increasing Cas9-mediated homology-directed repair efficiency through covalent tethering of DNA repair template的研究,描述了美国明尼苏达大学基因工程中心的Wendy Gordon的团队发现的能够增加Cas9介导的同源定向DNA修复的效率的研究。

近日在《通讯-生物学》发表的一项名为Increasing Cas9-mediated homology-directed repair efficiency through covalent tethering of DNA repair template的研究,描述了美国明尼苏达大学基因工程中心的Wendy Gordon的团队发现的能够增加Cas9介导的同源定向DNA修复的效率的研究。

CRISPR/Cas9是有力的基因组编辑工具。研究者们已经成功地找出用CRISPR/Cas9系统把基因关闭、启动或者修复的方法。但是因为CRISPR/Cas9系统的准确率不稳定,所以专家们在把其应用在众多疾病,比如X染色体断裂症候群,镰状细胞贫血,亨廷顿疾病的临床治疗上一直持保守态度。

研究者们也发现,不同的引导DNA,不同的靶基因,基因组里不同的区域,不同的细胞系,或者取自不同器官、组织、或者病人的细胞里,CRISPR/Cas9系统的成功率也大有不同。虽然有些环境会让CRISPR/Cas9的成功率高达近100%,但是多数的环境会导致很低甚至近于零的成功率。所以研究者们已经着手研究增加CRISPR/Cas9的成功率的策略,比如改变Cas9蛋白质的结构,加入其他的DNA片段来干扰细胞原有的DNA修复功能等等。但是这些策略仍然会受细胞种类或者靶基因在基因组里的位置的影响。



Cas9-PCV上ssDNA的选择性共价键键合

在现有的系统里,虽然Cas9蛋白质和引导DNA通常是被包装在一起送入靶细胞,但是它们之间没有共价键。以前已经有研究团队显示,把引导DNA和一个与Cas9相似的被改进的蛋白质用共价键结合,会增加其同源定向DNA的修复的效率。但是那个方法的工序过于复杂和昂贵。

于是Wendy Gordon的团队想寻找另外一个把引导DNA用共价键和Cas9结合,以增加CRISPR/Cas9的系统的效率的策略。他们之前发现,HUH内切酶和单链DNA之间能够容易并快速地形成磷酸酪氨酸键。于是他们设计和制造了一个HUH内切酶和Cas9的融合蛋白,然后证明这个融合蛋白能够和单链DNA形成共价键。他们的研究显示,这个融合蛋白和也是单链的引导DNA的复合体能够把同源定向修复的成功率增加30倍。他们的初步数据显示,这个策略不会受到细胞种类或靶基因在基因组里的位置的影响。而且这个方法比之前的方法更简洁和低成本。

Wendy Gordon的团队希望以这个策略为增加CRISPR/Cas9系统的效率做出贡献。

原始出处:
Eric J. Aird, Klaus N. Lovendahl, Amber St. Martin, et al. Increasing Cas9-mediated homology-directed repair efficiency through covalent tethering of DNA repair template. Communications Biology. 31 May 2018.

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    2018-08-30 sunylz
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    2019-04-24 liuli5079
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encryptionId=755d5211a0, topicName=CRISPR)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4c37265, createdName=hb2008ye, createdTime=Mon Aug 27 01:43:00 CST 2018, time=2018-08-27, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1341258, encodeId=ba411341258e7, content=<a href='/topic/show?id=e9c45226b3' target=_blank style='color:#2F92EE;'>#CRISPR/Cas9#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=46, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=5226, encryptionId=e9c45226b3, topicName=CRISPR/Cas9)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=7eeb2500172, createdName=haouestc, createdTime=Mon Aug 27 01:43:00 CST 2018, time=2018-08-27, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1365057, encodeId=013d136505e6f, content=<a href='/topic/show?id=3f66412ebd' target=_blank style='color:#2F92EE;'>#Cas9#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=22, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4127, encryptionId=3f66412ebd, topicName=Cas9)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=8346212, createdName=respect, createdTime=Mon Aug 27 01:43:00 CST 2018, time=2018-08-27, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1624887, encodeId=bdaf162488ed0, content=<a href='/topic/show?id=0333412062' target=_blank style='color:#2F92EE;'>#Cas#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=24, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4120, encryptionId=0333412062, topicName=Cas)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=411420689333, createdName=yaanren, createdTime=Mon Aug 27 01:43:00 CST 2018, time=2018-08-27, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1625774, encodeId=7bde1625e744e, content=<a href='/topic/show?id=4d072999181' target=_blank style='color:#2F92EE;'>#共价键#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=29991, encryptionId=4d072999181, topicName=共价键)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=276c20872742, createdName=millore, createdTime=Mon Aug 27 01:43:00 CST 2018, time=2018-08-27, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=340746, encodeId=3519340e46c3, content=好, beContent=null, objectType=article, channel=null, level=null, likeNumber=53, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=0aa31415219, createdName=orangesking, createdTime=Sat Aug 25 12:13:24 CST 2018, time=2018-08-25, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=340745, encodeId=7da0340e45a9, content=学习一下知识, beContent=null, objectType=article, channel=null, level=null, likeNumber=71, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://cacheapi.medsci.cn/resource/upload/20160718/IMG578C993D395567540.jpg, createdBy=59b81918946, createdName=flysky120, createdTime=Sat Aug 25 12:04:40 CST 2018, time=2018-08-25, status=1, ipAttribution=)]
    2018-08-27 hb2008ye
  5. 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content=<a href='/topic/show?id=4d072999181' target=_blank style='color:#2F92EE;'>#共价键#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=29991, encryptionId=4d072999181, topicName=共价键)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=276c20872742, createdName=millore, createdTime=Mon Aug 27 01:43:00 CST 2018, time=2018-08-27, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=340746, encodeId=3519340e46c3, content=好, beContent=null, objectType=article, channel=null, level=null, likeNumber=53, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=0aa31415219, createdName=orangesking, createdTime=Sat Aug 25 12:13:24 CST 2018, time=2018-08-25, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=340745, encodeId=7da0340e45a9, content=学习一下知识, beContent=null, objectType=article, channel=null, level=null, likeNumber=71, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://cacheapi.medsci.cn/resource/upload/20160718/IMG578C993D395567540.jpg, createdBy=59b81918946, createdName=flysky120, createdTime=Sat Aug 25 12:04:40 CST 2018, time=2018-08-25, status=1, ipAttribution=)]
  6. [GetPortalCommentsPageByObjectIdResponse(id=1950132, encodeId=474b195013210, content=<a href='/topic/show?id=f98b15504d8' target=_blank style='color:#2F92EE;'>#RISPR/Cas9#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=42, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=15504, encryptionId=f98b15504d8, topicName=RISPR/Cas9)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=e1cc90, createdName=rebeccajiejie, createdTime=Wed Jan 16 04:43:00 CST 2019, time=2019-01-16, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1918258, encodeId=f4361918258cb, content=<a href='/topic/show?id=4f26341994' target=_blank style='color:#2F92EE;'>#Bio#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=30, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=3419, encryptionId=4f26341994, topicName=Bio)], attachment=null, authenticateStatus=null, createdAvatar=, 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objectType=article, channel=null, level=null, likeNumber=22, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4127, encryptionId=3f66412ebd, topicName=Cas9)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=8346212, createdName=respect, createdTime=Mon Aug 27 01:43:00 CST 2018, time=2018-08-27, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1624887, encodeId=bdaf162488ed0, content=<a href='/topic/show?id=0333412062' target=_blank style='color:#2F92EE;'>#Cas#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=24, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4120, encryptionId=0333412062, topicName=Cas)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=411420689333, createdName=yaanren, createdTime=Mon Aug 27 01:43:00 CST 2018, time=2018-08-27, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1625774, encodeId=7bde1625e744e, content=<a href='/topic/show?id=4d072999181' target=_blank style='color:#2F92EE;'>#共价键#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=29991, encryptionId=4d072999181, topicName=共价键)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=276c20872742, createdName=millore, createdTime=Mon Aug 27 01:43:00 CST 2018, time=2018-08-27, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=340746, encodeId=3519340e46c3, content=好, beContent=null, objectType=article, channel=null, level=null, likeNumber=53, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=0aa31415219, createdName=orangesking, createdTime=Sat Aug 25 12:13:24 CST 2018, time=2018-08-25, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=340745, encodeId=7da0340e45a9, content=学习一下知识, beContent=null, objectType=article, channel=null, level=null, likeNumber=71, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://cacheapi.medsci.cn/resource/upload/20160718/IMG578C993D395567540.jpg, createdBy=59b81918946, createdName=flysky120, createdTime=Sat Aug 25 12:04:40 CST 2018, time=2018-08-25, status=1, ipAttribution=)]
    2018-08-27 respect
  7. 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objectType=article, channel=null, level=null, likeNumber=22, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4127, encryptionId=3f66412ebd, topicName=Cas9)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=8346212, createdName=respect, createdTime=Mon Aug 27 01:43:00 CST 2018, time=2018-08-27, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1624887, encodeId=bdaf162488ed0, content=<a href='/topic/show?id=0333412062' target=_blank style='color:#2F92EE;'>#Cas#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=24, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4120, encryptionId=0333412062, topicName=Cas)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=411420689333, createdName=yaanren, createdTime=Mon Aug 27 01:43:00 CST 2018, time=2018-08-27, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1625774, encodeId=7bde1625e744e, content=<a href='/topic/show?id=4d072999181' target=_blank style='color:#2F92EE;'>#共价键#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=29991, encryptionId=4d072999181, topicName=共价键)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=276c20872742, createdName=millore, createdTime=Mon Aug 27 01:43:00 CST 2018, time=2018-08-27, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=340746, encodeId=3519340e46c3, content=好, beContent=null, objectType=article, channel=null, level=null, likeNumber=53, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=0aa31415219, createdName=orangesking, createdTime=Sat Aug 25 12:13:24 CST 2018, time=2018-08-25, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=340745, encodeId=7da0340e45a9, content=学习一下知识, beContent=null, objectType=article, channel=null, level=null, likeNumber=71, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://cacheapi.medsci.cn/resource/upload/20160718/IMG578C993D395567540.jpg, createdBy=59b81918946, createdName=flysky120, createdTime=Sat Aug 25 12:04:40 CST 2018, time=2018-08-25, status=1, ipAttribution=)]
    2018-08-27 yaanren
  8. 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objectType=article, channel=null, level=null, likeNumber=71, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://cacheapi.medsci.cn/resource/upload/20160718/IMG578C993D395567540.jpg, createdBy=59b81918946, createdName=flysky120, createdTime=Sat Aug 25 12:04:40 CST 2018, time=2018-08-25, status=1, ipAttribution=)]
    2018-08-27 millore
  9. 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objectType=article, channel=null, level=null, likeNumber=71, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://cacheapi.medsci.cn/resource/upload/20160718/IMG578C993D395567540.jpg, createdBy=59b81918946, createdName=flysky120, createdTime=Sat Aug 25 12:04:40 CST 2018, time=2018-08-25, status=1, ipAttribution=)]
    2018-08-25 orangesking

    0

  10. 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    2018-08-25 flysky120

    学习一下知识

    0

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