优化基于ABA的化学诱导的近距离,以增强细胞内转录激活和对ABA的修饰反应
Zeng Zhou1,2, Yue-Qi Wang3, Xu-Nan Zheng3
1College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002, China.
Science China. Life sciences
|August 22, 2024
概括
使用酸 (ABA) 的化学诱导近距离 (CIP) 通过设计PYR1-ABI1相互作用来增强,从而改善了转录激活和RNA修饰以抑制瘤细胞增殖.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 合成生物学 合成生物学
背景情况:
- 基于酸 (ABA) 的化学诱导近距离 (CIP) 系统依赖于ABA受体 (如PYL1) 和蛋白质酸酶 (如ABI1) 之间的相互作用.
- 目前基于ABA的CIP应用程序受到亚最佳ABA响应强度的限制.
- 提高ABA诱导的蛋白质与蛋白质相互作用的强度对于更广泛的应用至关重要.
研究的目的:
- 开发和增强基于ABA的CIP系统,以提高响应强度.
- 为哺乳动物细胞设计ABA诱导的转录激活工具.
- 为了探索治疗应用的ABA诱导的RNA m6A修饰.
主要方法:
- 在HEK293T细胞中对PYR1-ABI1和PYL1-ABI1之间的ABA诱导相互作用进行比较分析.
- 基于CRISPR/dCas9的转录激活工具的工程使用PYR1-ABI1和PYL1-ABI1.1.
- 用ABA诱导的PYR1-ABI1相互作用与CRISPR/dCas13用于RNAm6A修饰的应用.
- 利用菌体辅助连续进化 (PACE) 来改善PYR1-ABI1的相互作用.
主要成果:
- 与PYL1-ABI1.1相比,PYR1-ABI1相互作用显示出较高的ABA反应强度.
- 基于PYR1-ABI1的转录激活工具对外源和内源基因都表现出卓越的性能.
- 使用PYR1-ABI1-CRISPR/dCas13的ABA诱导的RNA m6A修饰成功抑制了瘤细胞的增殖.
- PACE产生了一种PYR1突变 (PYR1m) 与ABI1的相互作用强度增强,进一步改进了转录激活工具.
结论:
- 开发了一个基于ABA的新型CIP系统,增强了响应强度.
- PYR1-ABI1相互作用作为一个更强大的模块,用于ABA诱导的近距离应用.
- PACE是优化CIP系统的有效策略.
- 这种改进的ABA-CIP方法为修改生物过程和潜在的治疗干预提供了新的途径.
相关概念视频
Chromatin Modification in iPS Cells
1.6K
Chromatin modification alters gene expression; therefore, scientists can add histone-modifying enzymes, histone variants, and chromatin remodeling complexes to somatic cells to aid reprogramming into pluripotent stem (iPS) cells.
Compact chromatin makes reprogramming difficult. Enzymes, such as histone demethylases and acetyltransferases, are often added during reprogramming to loosen the chromatin, making the DNA more accessible to transcription factors. Molecules that inhibit histone...
Compact chromatin makes reprogramming difficult. Enzymes, such as histone demethylases and acetyltransferases, are often added during reprogramming to loosen the chromatin, making the DNA more accessible to transcription factors. Molecules that inhibit histone...
1.6K
RNA Polymerase II Accessory Proteins
9.1K
Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...
9.1K
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
Eukaryotic Transcription Activators
10.9K
Transcription activators are proteins that promote the transcription of genes from DNA to RNA. In most cases, these proteins contain two separate domains ‒ a domain that binds to DNA and a domain for activating transcription; however, in some cases, a single domain is responsible for both binding and activation of transcription, as seen in the glucocorticoid receptor and MyoD.
The binding domains are capable of recognizing and interacting with regulatory sequences on the DNA. These...
The binding domains are capable of recognizing and interacting with regulatory sequences on the DNA. These...
10.9K
Covalently Linked Protein Regulators
6.8K
Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein....
These groups modify specific amino acids in a protein....
6.8K
Transcription Attenuation in Prokaryotes
15.2K
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...
15.2K


