南方科技大学知识苑(SUSTech KC): THE FUNCTIONAL STUDY OF PROTEIN INTRINSIC DISORDER REGIONS IN CRISPR MEDIATED GENE TRANSCRIPTIONAL REGULATION

题名	THE FUNCTIONAL STUDY OF PROTEIN INTRINSIC DISORDER REGIONS IN CRISPR MEDIATED GENE TRANSCRIPTIONAL REGULATION
其他题名	蛋白内在无序区在 CRISPR 介导的基因转录调控中的功能作用
姓名	史心瑶
姓名拼音	SHI Xinyao
学号	12032181
学位类型	硕士
学位专业	071007 遗传学
学科门类/专业学位类别	07 理学
导师	崔欢欢
导师单位	系统生物学系
论文答辩日期	2023-05-15
论文提交日期	2023-06-30
学位授予单位	南方科技大学
学位授予地点	深圳
摘要	CRISPR mediated transcription regulation system is based on an RNA-guided inactive Cas9 (dCas9) acting as a programmable transcription regulator via fusing effector domains. However, inefficiencies in target gene activation or repression have limited its potential applications. Recent studies have shown that transcription factors (TFs) contain intrinsic disorder regions (IDRs) in their activation domains, and the multivalent interactions between IDRs play an important role in transcription activation. Nevertheless, the causal association between IDRs and transcription regulation remains unclear. Here, we combined IDR with CRISPR mediated transcription regulation system to further improve its efficiency and tried to uncover the association between IDRs and transcription regulation. By fusing IDR with CRISPR activation system dCas9- VP64, we demonstrated that dCas9-VP64-IDR functioned as an enhanced dCas9 activator with the improved activation efficiency being attributed to the multivalent interaction between IDRs. However, not all IDRs can exceptionally amplify the activation signal. Moreover, when we fused IDR with the CRISPR repression system dCas9-KRAB, we observed almost no alterations of repression efficiency on endogenous genes. Further living cell images analysis indicated that the activation efficiency and the capacity to form phase separation are not necessarily correlated, suggesting that the improvement of transcription efficiency is independent of phase separation. Taken together, our findings reveal that IDR-mediated multivalent interactions can enhance gene activation, and that our dCas9-AD-IDR can substantially improve the efficiency of the current CRISPR activation system, but not for CRISPR mediated gene repression, and we partly uncover the relationship between IDRs and transcription regulation. These results together may contribute to the studies on the relationship between phase separation and transcriptional regulation.
关键词	CRISPR/Cas9 IDR Transcriptional Regulation Multivalent Interaction Phase Separation
语种	英语
培养类别	独立培养
入学年份	2020
学位授予年份	2023-06
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所在学位评定分委会	生物学
国内图书分类号	Q754
来源库	人工提交
成果类型	学位论文
条目标识符	http://sustech.caswiz.com/handle/2SGJ60CL/544745
专题	生命科学学院_生物系
推荐引用方式 GB/T 7714	Shi XY. THE FUNCTIONAL STUDY OF PROTEIN INTRINSIC DISORDER REGIONS IN CRISPR MEDIATED GENE TRANSCRIPTIONAL REGULATION[D]. 深圳. 南方科技大学,2023.

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