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Science:科学家发现新型基因编辑工具,体积是CRISPR-Cas9的一半

2020-07-22 佚名 探索菌

基因编辑作为一种新兴的基因工程技术,其本质是人为地改变自然选择。目前广泛使用的“基因剪刀”是CRISPR-Cas9,该技术在一系列基因治疗的应用领域都展现出极大的应用前景。

基因编辑作为一种新兴的基因工程技术,其本质是人为地改变自然选择。目前广泛使用的“基因剪刀”是CRISPR-Cas9,该技术在一系列基因治疗的应用领域都展现出极大的应用前景,但过大的蛋白分子成为CRISPR未来应用方式中最重要的瓶颈之一。


最近,来自美国加利福尼亚大学伯克利分校创新基因组学研究所的研究人员发现了一种新型基因编辑工具——CasΦ,它是一种功能最小的CRISPR-Cas系统,由1?70 KD的CasΦ蛋白和一个CRISPR阵列组成,且仅在巨型噬菌体的基因组中编码,其体积是CRISPR-Cas 9的一半,这项研究成果于7月16日发表在《Science》杂志上。


CasΦ(Cas12j)是巨型噬菌体分枝中编码的Cas蛋白家族,巨型噬菌体用它来诱使细菌抵抗自身而不是自身的病毒。它使用单个活性位点进行CRISPR RNA(crRNA)加工和crRNA指导的DNA切割,以靶向外来核酸。


研究人员首先要确定CasΦ是否能作为真正的CRISPR-Cas系统。他们发现,CasΦ在其天然环境中是一种功能性噬菌体蛋白和真正的CRISPR-Cas效应子,能够裂解crRNA互补的DNA。此外,这种单RNA系统比其他活性CRISPR-Cas系统紧凑得多。


接下来,研究小组需要了解CasΦ在体外对DNA的识别和裂解要求。结果表明,CasΦ的裂解活性仅在识别顺式DNA时才能观察到,而且只需要最低的PAM条件,这可能有利于更广泛的核酸检测。


最后,为了研究CasΦ是否能用于人类基因组编辑,研究人员在HEK293细胞中使用了与crRNA共表达的CasΦ进行了基因破坏测定。他们发现,CasΦ-2和CasΦ-3诱导了基因组整合增强型绿色荧光蛋白(EGFP)基因的定向破坏。其中,具有单个指导RNA的CasΦ-2能够编辑多达33%的细胞,这与先前报道的CRISPR-Cas9,CRISPR-Cas12a和CRISPR-CasX的水平相当。


其实,这并不是CasΦ的首次亮相。它最早是由这项研究的另一位主要作者——IGI的一名博士生Basem Al-Shayeb发现的,这项研究于今年2月12日发表在《Nature》上。当时,Al-Shayeb正在加州大学伯克利分校地球与行星科学系教授Jillian F. Banfield的实验室工作。他们认为,这种含有CasΦ的巨型噬菌体是已有巨型噬菌体的成员之一,并且存在于多种环境中。


最近的这一发现揭示了CasΦ巨大的基因编辑潜力,从而为细胞传递提供了优势,扩大了基因编辑的“工具箱”。更重要的是,CasΦ蛋白将巨型噬菌体推向人类健康发现和生物技术应用的前沿。但对于优化CasΦ用于基因编辑,以及探索用于设计靶向特定基因的指导RNA的最佳方法上,研究人员还有很长的路要走。

 

原始出处:

Patrick Pausch , Basem Al-Shayeb  Ezra Bisom-Rapp , et al.CRISPR-CasΦ from huge phages is a hypercompact genome editor.Science. 2020 Jul 17;369(6501):333-337. doi:10.1126/science.abb1400.

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    2020-07-28 ms 内分泌科张

    0

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    2020-07-24 yuandd
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  6. [GetPortalCommentsPageByObjectIdResponse(id=1802651, encodeId=07531802651c5, content=<a href='/topic/show?id=acfae485578' target=_blank style='color:#2F92EE;'>#科学家发现#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=45, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=74855, encryptionId=acfae485578, topicName=科学家发现)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=e572480, createdName=旅苦化文_208, createdTime=Sat Nov 07 21:44:28 CST 2020, time=2020-11-07, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1994057, encodeId=773c199405e20, content=<a href='/topic/show?id=3b1d4186600' target=_blank style='color:#2F92EE;'>#基因编辑工具#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=66, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=41866, encryptionId=3b1d4186600, topicName=基因编辑工具)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=322359, createdName=bioon3, createdTime=Sat Feb 27 08:44:28 CST 2021, time=2021-02-27, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=804922, encodeId=3ae680492296, content=。, beContent=null, objectType=article, channel=null, level=null, likeNumber=109, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://img.medsci.cn/20201214/b3170121d995432ea306ae3a7809cc05/2cfb236f361b4d3c80ef8fdef4301832.jpg, createdBy=8ad35400531, createdName=ms 内分泌科张, createdTime=Tue Jul 28 18:50:46 CST 2020, time=2020-07-28, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1276161, encodeId=bdf112e6161bc, content=<a href='/topic/show?id=755d5211a0' target=_blank style='color:#2F92EE;'>#CRISPR#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=47, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=5211, encryptionId=755d5211a0, topicName=CRISPR)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=583c133, createdName=yuandd, createdTime=Fri Jul 24 08:44:28 CST 2020, time=2020-07-24, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1360450, encodeId=be4a1360450a6, content=<a href='/topic/show?id=99e5521532' target=_blank style='color:#2F92EE;'>#CRISPR-Cas9#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=59, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=5215, encryptionId=99e5521532, topicName=CRISPR-Cas9)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=28b7142, createdName=liuquan, createdTime=Fri Jul 24 08:44:28 CST 2020, time=2020-07-24, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1365074, encodeId=4cdb13650e438, content=<a href='/topic/show?id=3f66412ebd' target=_blank style='color:#2F92EE;'>#Cas9#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=57, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4127, encryptionId=3f66412ebd, topicName=Cas9)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=8346212, createdName=respect, createdTime=Fri Jul 24 08:44:28 CST 2020, time=2020-07-24, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1376133, encodeId=bfd713e61335d, content=<a href='/topic/show?id=2949159989d' target=_blank style='color:#2F92EE;'>#SCIE#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=42, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=15998, encryptionId=2949159989d, topicName=SCIE)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=17a3387, createdName=jichang, createdTime=Fri Jul 24 08:44:28 CST 2020, time=2020-07-24, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1455293, encodeId=901214552937c, content=<a href='/topic/show?id=ce8452137f' target=_blank style='color:#2F92EE;'>#CRISPR-Cas#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=60, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=5213, encryptionId=ce8452137f, topicName=CRISPR-Cas)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=e8505649833, createdName=guihongzh, createdTime=Fri Jul 24 08:44:28 CST 2020, time=2020-07-24, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1624957, encodeId=cd14162495eaa, content=<a href='/topic/show?id=0333412062' target=_blank style='color:#2F92EE;'>#Cas#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=52, 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=Fri Jul 24 08:44:28 CST 2020, time=2020-07-24, status=1, ipAttribution=)]
    2020-07-24 respect
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    2020-07-24 jichang
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    2020-07-24 yaanren

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