通过TPR-CHAT对CRISPR-Cas7-11进行负调节的结构基础
Tian Hong1, Qinghua Luo1,2, Haiyun Ma1
1Department of Laboratory Medicine, State Key Laboratory of Biotherapy, National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University, Chengdu, China.
Signal transduction and targeted therapy
|May 12, 2024
概括
该CRISPR-Cas7-11RNA编辑工具由TPR-CHAT进行调节. TPR-CHAT的N终端域抑制了Cas7-11的活动,提高了CRISPR系统的安全性.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 结构生物学 结构生物学
背景情况:
- CRISPR-Cas7-11是一种用于RNA编辑的III-E型CRISPR相关核酶.
- TPR-CHAT是一种调节蛋白,它与Cas7-11.一起形成CRISPR引导的酶复合体 (Craspase).
研究的目的:
- 研究TPR-CHAT对Cas7-11核酶活性调节的结构基础.
- 了解克拉斯帕斯复合物的功能以及TPR-CHAT如何调节Cas7-11.
主要方法:
- 电子显微镜 (cryo-EM) 用于确定与TPR-CHAT变体复杂的DiCas7-11-crRNA的结构.
- 试验室内核酶测定测量Cas7-11活动.
- 电泳运动转移试验 (EMSA) 来评估向RNA的结合.
主要成果:
- 冷电磁结构揭示了克拉斯帕斯复合体的分子结构.
- 通过抑制标RNA结合,DiTPR-CHAT通过抑制标RNA结合来负面调节DiCas7-11的活动.
- DiTPR-CHAT (DiTPR-CHATNTD) 的N端65个氨基酸足以进行这种抑制功能.
结论:
- DiTPR-CHATNTD 作为 DiCas7-11.11 的最小监管单位.
- 这一发现表明,通过控制核酶活性和减少非目标效应,有可能开发更安全的CRISPR-Cas7-11应用.
相关概念视频
CRISPR and crRNAs
17.0K
Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
17.0K
CRISPR
50.7K
Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
50.7K
Transcription Attenuation in Prokaryotes
15.3K
Transcriptional attenuation occurs when RNA transcription is prematurely terminated due to the formation of a terminator mRNA hairpin structure. Bacteria use these hairpins to regulate the transcription process and control the synthesis of several amino acids including histidine, lysine, threonine, and phenylalanine. Transcription attenuation takes place in the non-coding regions of mRNA.
There are several different mechanisms used to attenuate transcription. In ribosome mediated...
There are several different mechanisms used to attenuate transcription. In ribosome mediated...
15.3K
Negative Regulator Molecules
35.3K
Positive regulators allow a cell to advance through cell cycle checkpoints. Negative regulators have an equally important role as they terminate a cell’s progression through the cell cycle—or pause it—until the cell meets specific criteria.
35.3K
Co-activators and Co-repressors
7.3K
Gene transcription is regulated by the synergistic action of several proteins that form a complex at a gene regulatory site. This is observed in eukaryotes, where the regulation of gene expression is a complex process. Regulatory proteins in eukaryotes can broadly be classified into two types – regulators that bind directly to specific DNA sequences and co-regulators that associate with regulatory proteins but cannot directly bind to the DNA. These co-regulators are further divided into...
7.3K
Cis-regulatory Sequences
9.9K
Cis-regulatory sequences are short fragments of non-coding DNA that are present on the same chromosomes as the genes that they regulate. These fragments serve as binding sites for transcriptional regulators, proteins that are responsible for controlling gene transcription and differential gene expression across cell types in eukaryotes. Cis-regulatory sequences can be close to the gene of interest or thousands of bases away in the DNA sequence; however, those sequences that are further away are...
9.9K


