Protocol to alter a protein's phase separation capacity to control cell fate transitions.

Protocol to alter a protein's phase separation capacity to control cell fate transitions.
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改变蛋白质相分离能力以控制细胞命运转变的方案

DOI:
10.1016/j.xpro.2021.100887
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发表时间:
2021-12-17
期刊:
影响因子:
--
通讯作者:
Ding J
Ding J
中科院分区:
其他
文献类型:
--
作者:
Ma Q;Wang J;Yu H;Sun J;Sun J;Chen J;Yu Y;Fang P;Tian Q;Ding J

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蛋白质的相分离调节转录。在这里,我们提出了一个协议来操纵相分离能力的蛋白质。我们使用这个协议来破坏相分离的突变残基在本质无序区(IDRs)。此外,我们通过融合已知的驱动相分离的IDR来挽救被禁用的相分离。相分离促进细胞命运的转变,而相破坏则减弱这种转变。主要的挑战是如何有效地预测突变残基。有关使用和执行本协议的完整详情,请参阅。在体外和体内均可验证具有液体样行为的相分离,IDR的残基突变可破坏蛋白质的相分离能力,通过融合IDR可恢复被破坏的蛋白质相分离,相分离调节细胞命运转变,相分离调节转录。在这里,我们提出了一个协议来操纵相分离能力的蛋白质。我们使用这个协议来破坏相分离的突变残基在本质无序区(IDRs)。此外,我们通过融合已知的驱动相分离的IDR来挽救被禁用的相分离。相分离促进细胞命运的转变,而相破坏则减弱这种转变。主要的挑战是如何有效地预测突变残基。
Phase separation of proteins regulates transcription. Here, we present a protocol to manipulate phase separation capacity of a protein. We use this protocol to disrupt phase separation by mutating residues at intrinsically disordered regions (IDRs). Further, we rescue the disabled phase separation by fusing an IDR known to drive phase separation. Phase separation promotes cell fate transitions, whereas disruption of phase attenuates the transitions. The major challenge is how to effectively predict mutation residues. For complete details on the use and execution of this protocol, please refer to. Phase separation with liquid-like behavior can be verified in vitro and in vivo Residue mutation at IDR can disrupt phase separation capacity of a protein The disabled phase separation of a protein can be rescued by fusing an IDR Phase separation regulates cell fate transitions Phase separation of proteins regulates transcription. Here, we present a protocol to manipulate phase separation capacity of a protein. We use this protocol to disrupt phase separation by mutating residues at intrinsically disordered regions (IDRs). Further, we rescue the disabled phase separation by fusing an IDR known to drive phase separation. Phase separation promotes cell fate transitions, whereas disruption of phase attenuates the transitions. The major challenge is how to effectively predict mutation residues.
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