RNA Biomedical Institute, Sun Yat-Sen Memorial Hospital, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou 510080, China.
Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-Sen University, Guangzhou 510080, China.
STAR Protoc. 2021 Oct 14;2(4):100887. doi: 10.1016/j.xpro.2021.100887. eCollection 2021 Dec 17.
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 Wang et al. (2021).
蛋白质的液-液相分离调控转录。在此，我们提供了一种操纵蛋白质液-液相分离能力的方案。我们使用该方案通过突变无序区域（IDR）的残基来破坏液-液相分离。此外，我们通过融合已知能驱动液-相分离的 IDR 来挽救失活的液-相分离。液-相分离促进细胞命运转变，而破坏液-相分离则会减弱转变。主要的挑战是如何有效地预测突变残基。有关该方案使用和执行的完整详细信息，请参考 Wang 等人（2021 年）。
