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Methods Mol Biol:利用CRISPR / Cas9构建视网膜变性模型

2018-11-12 MedSci MedSci原创

加拿大哥伦比亚大学眼科学与视觉科学系的Feehan JM等人近日在Methods Mol Biol杂志上发表了一篇文章,他们利用CRISPR / Cas9基因编辑技术构建了视网膜变性的动物模型。

加拿大哥伦比亚大学眼科学与视觉科学系的Feehan JM等人近日在Methods Mol Biol杂志上发表了一篇文章,他们利用CRISPR / Cas9基因编辑技术构建了视网膜变性的动物模型。

之前的研究已证明非洲爪蟾(Xenopus laevis)是可以用于快速构建和分析人视网膜疾病转基因模型的有效系统。然而,该系统缺乏稳健的基因插入和敲除的试验技术,因此受到一定的限制。在这里,他们通过使用Cas9基因编辑技术,构建了X.laevis胚胎,并对这个方案进行了详细的描述。

他们使用该技术将点突变引入X.laevis的基因组中,使阅读框内和框外产生插入和缺失。构建的动物模型允许对人类显性和隐性遗传疾病进行建模,并可以有效地产生基因敲低和敲除的效果。

因此,他们认为,他们的技术可以在X. laevis中进行高频基因编辑,同时可以在F0代中进行表型分析。

原文出处:

Feehan, J.M., et al., Generation and Analysis of Xenopus laevis Models of Retinal Degeneration Using CRISPR/Cas9. Methods Mol Biol, 2019. 1834: p. 193-207.

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status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1463645, encodeId=fc43146364525, content=<a href='/topic/show?id=7d27381288e' target=_blank style='color:#2F92EE;'>#变性#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=57, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=38128, encryptionId=7d27381288e, topicName=变性)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=f69f6545041, createdName=huangdf, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=)]
  5. [GetPortalCommentsPageByObjectIdResponse(id=1896418, encodeId=5684189641846, 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=Sat May 18 21:24:00 CST 2019, time=2019-05-18, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1747101, encodeId=53ba1e471010f, content=<a href='/topic/show?id=76a090e5544' target=_blank style='color:#2F92EE;'>#视网膜变性#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=72, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=90755, encryptionId=76a090e5544, topicName=视网膜变性)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=a6db35920976, 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status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1463645, encodeId=fc43146364525, content=<a href='/topic/show?id=7d27381288e' target=_blank style='color:#2F92EE;'>#变性#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=57, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=38128, encryptionId=7d27381288e, topicName=变性)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=f69f6545041, createdName=huangdf, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=)]
  6. [GetPortalCommentsPageByObjectIdResponse(id=1896418, encodeId=5684189641846, 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=Sat May 18 21:24:00 CST 2019, time=2019-05-18, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1747101, encodeId=53ba1e471010f, content=<a href='/topic/show?id=76a090e5544' target=_blank style='color:#2F92EE;'>#视网膜变性#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=72, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=90755, encryptionId=76a090e5544, topicName=视网膜变性)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=a6db35920976, 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beContent=null, objectType=article, channel=null, level=null, likeNumber=56, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=3419, encryptionId=4f26341994, topicName=Bio)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=12de429, createdName=sunylz, createdTime=Mon Apr 15 15:24:00 CST 2019, time=2019-04-15, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1270132, encodeId=171112e013293, content=<a href='/topic/show?id=979690e33bd' target=_blank style='color:#2F92EE;'>#视网膜#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=90, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=90733, encryptionId=979690e33bd, topicName=视网膜)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=742793, createdName=zutt, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1276102, encodeId=f96412e610274, content=<a href='/topic/show?id=755d5211a0' target=_blank style='color:#2F92EE;'>#CRISPR#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=57, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=5211, encryptionId=755d5211a0, topicName=CRISPR)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=583c133, createdName=yuandd, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1365060, encodeId=ab5d1365060fd, content=<a href='/topic/show?id=3f66412ebd' target=_blank style='color:#2F92EE;'>#Cas9#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=61, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4127, encryptionId=3f66412ebd, topicName=Cas9)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=8346212, createdName=respect, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1463645, encodeId=fc43146364525, content=<a href='/topic/show?id=7d27381288e' target=_blank style='color:#2F92EE;'>#变性#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=57, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=38128, encryptionId=7d27381288e, topicName=变性)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=f69f6545041, createdName=huangdf, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=)]
    2019-04-15 sunylz
  7. 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beContent=null, objectType=article, channel=null, level=null, likeNumber=56, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=3419, encryptionId=4f26341994, topicName=Bio)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=12de429, createdName=sunylz, createdTime=Mon Apr 15 15:24:00 CST 2019, time=2019-04-15, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1270132, encodeId=171112e013293, content=<a href='/topic/show?id=979690e33bd' target=_blank style='color:#2F92EE;'>#视网膜#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=90, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=90733, encryptionId=979690e33bd, topicName=视网膜)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=742793, createdName=zutt, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1276102, encodeId=f96412e610274, content=<a href='/topic/show?id=755d5211a0' target=_blank style='color:#2F92EE;'>#CRISPR#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=57, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=5211, encryptionId=755d5211a0, topicName=CRISPR)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=583c133, createdName=yuandd, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1365060, encodeId=ab5d1365060fd, content=<a href='/topic/show?id=3f66412ebd' target=_blank style='color:#2F92EE;'>#Cas9#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=61, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4127, encryptionId=3f66412ebd, topicName=Cas9)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=8346212, createdName=respect, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1463645, encodeId=fc43146364525, content=<a href='/topic/show?id=7d27381288e' target=_blank style='color:#2F92EE;'>#变性#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=57, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=38128, encryptionId=7d27381288e, topicName=变性)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=f69f6545041, createdName=huangdf, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=)]
    2018-11-14 zutt
  8. [GetPortalCommentsPageByObjectIdResponse(id=1896418, encodeId=5684189641846, 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=Sat May 18 21:24:00 CST 2019, time=2019-05-18, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1747101, encodeId=53ba1e471010f, content=<a href='/topic/show?id=76a090e5544' target=_blank style='color:#2F92EE;'>#视网膜变性#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=72, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=90755, encryptionId=76a090e5544, topicName=视网膜变性)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=a6db35920976, 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beContent=null, objectType=article, channel=null, level=null, likeNumber=56, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=3419, encryptionId=4f26341994, topicName=Bio)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=12de429, createdName=sunylz, createdTime=Mon Apr 15 15:24:00 CST 2019, time=2019-04-15, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1270132, encodeId=171112e013293, content=<a href='/topic/show?id=979690e33bd' target=_blank style='color:#2F92EE;'>#视网膜#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=90, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=90733, encryptionId=979690e33bd, topicName=视网膜)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=742793, createdName=zutt, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1276102, encodeId=f96412e610274, content=<a href='/topic/show?id=755d5211a0' target=_blank style='color:#2F92EE;'>#CRISPR#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=57, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=5211, encryptionId=755d5211a0, topicName=CRISPR)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=583c133, createdName=yuandd, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1365060, encodeId=ab5d1365060fd, content=<a href='/topic/show?id=3f66412ebd' target=_blank style='color:#2F92EE;'>#Cas9#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=61, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4127, encryptionId=3f66412ebd, topicName=Cas9)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=8346212, createdName=respect, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1463645, encodeId=fc43146364525, content=<a href='/topic/show?id=7d27381288e' target=_blank style='color:#2F92EE;'>#变性#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=57, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=38128, encryptionId=7d27381288e, topicName=变性)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=f69f6545041, createdName=huangdf, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=)]
    2018-11-14 yuandd
  9. 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createdName=circumcision, createdTime=Mon Apr 15 04:24:00 CST 2019, time=2019-04-15, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1767557, encodeId=ce381e6755766, content=<a href='/topic/show?id=64f5343e2f' target=_blank style='color:#2F92EE;'>#Biol#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=68, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=3437, encryptionId=64f5343e2f, topicName=Biol)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=446937890434, createdName=12498ebem31暂无昵称, createdTime=Thu Jun 13 13:24:00 CST 2019, time=2019-06-13, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=2034052, encodeId=5aa220340525a, content=<a href='/topic/show?id=860752127a' target=_blank style='color:#2F92EE;'>#CRISPR / Cas9#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=68, replyNumber=0, topicName=null, topicId=null, 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beContent=null, objectType=article, channel=null, level=null, likeNumber=56, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=3419, encryptionId=4f26341994, topicName=Bio)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=12de429, createdName=sunylz, createdTime=Mon Apr 15 15:24:00 CST 2019, time=2019-04-15, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1270132, encodeId=171112e013293, content=<a href='/topic/show?id=979690e33bd' target=_blank style='color:#2F92EE;'>#视网膜#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=90, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=90733, encryptionId=979690e33bd, topicName=视网膜)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=742793, createdName=zutt, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1276102, encodeId=f96412e610274, content=<a href='/topic/show?id=755d5211a0' target=_blank style='color:#2F92EE;'>#CRISPR#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=57, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=5211, encryptionId=755d5211a0, topicName=CRISPR)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=583c133, createdName=yuandd, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1365060, encodeId=ab5d1365060fd, content=<a href='/topic/show?id=3f66412ebd' target=_blank style='color:#2F92EE;'>#Cas9#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=61, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4127, encryptionId=3f66412ebd, topicName=Cas9)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=8346212, createdName=respect, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1463645, encodeId=fc43146364525, content=<a href='/topic/show?id=7d27381288e' target=_blank style='color:#2F92EE;'>#变性#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=57, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=38128, encryptionId=7d27381288e, topicName=变性)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=f69f6545041, createdName=huangdf, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=)]
    2018-11-14 respect
  10. 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content=<a href='/topic/show?id=755d5211a0' target=_blank style='color:#2F92EE;'>#CRISPR#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=57, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=5211, encryptionId=755d5211a0, topicName=CRISPR)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=583c133, createdName=yuandd, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1365060, encodeId=ab5d1365060fd, content=<a href='/topic/show?id=3f66412ebd' target=_blank style='color:#2F92EE;'>#Cas9#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=61, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4127, encryptionId=3f66412ebd, topicName=Cas9)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=8346212, createdName=respect, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1463645, encodeId=fc43146364525, content=<a href='/topic/show?id=7d27381288e' target=_blank style='color:#2F92EE;'>#变性#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=57, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=38128, encryptionId=7d27381288e, topicName=变性)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=f69f6545041, createdName=huangdf, createdTime=Wed Nov 14 12:24:00 CST 2018, time=2018-11-14, status=1, ipAttribution=)]
    2018-11-14 huangdf

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