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相关概念视频

Caspases01:24

Caspases

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Caspase, a family of cysteine proteases, serve as effectors in apoptosis. The ced3 gene in C.elegans was first identified to be involved in apoptosis. This gene encodes the ced-3 caspase that is similar to the interleukin-1-beta converting enzyme or ICE in mammals. In addition to apoptosis, caspases also function in the inflammatory response. Inflammatory caspases are essential in activating pro-inflammatory cytokines that recruit immune cells and block the replication of pathogens inside...
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Regulation of Nuclear Protein Sorting01:45

Regulation of Nuclear Protein Sorting

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Nuclear protein sorting regulates nucleus composition and gene expression, crucial for determining the fate of a eukaryotic cell. Hence, the entry and exit of molecules across the nuclear envelope is a tightly controlled process. Nuclear protein sorting can be inhibited by one of the following ways: 1) masking cargo signal sequences, 2) modifying the nuclear receptor's affinity for cargo, 3) controlling the nuclear pore size, 4) retaining the cargo during its transit to the cytosol or the...
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CRISPR and crRNAs02:53

CRISPR and crRNAs

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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...
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Catenins01:23

Catenins

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Catenins are characterized by multiple binding domains and dynamic structures that allow them to function as linker proteins in cell junction complexes. All catenins, except α-catenin, contain a characteristic protein sequence called the armadillo repeat and are therefore also called armadillo proteins.
Catenins in Cell Junctions
Catenins bind to cell adhesion molecules such as cadherins and link them to different cytoskeletal proteins depending on the type of cell junction. At the...
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Protein Complexes with Interchangeable Parts01:57

Protein Complexes with Interchangeable Parts

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Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
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Assembly of Signaling Complexes

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Multiprotein signaling complexes are formed in a dynamic process involving protein-protein interactions at the cytoplasmic domain of transmembrane receptors or enzymatic and non-enzymatic proteins associated with the receptor. These complexes ensure the activation and propagation of intracellular signals that regulate cell functions.
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相关实验视频

Updated: Jul 1, 2025

Lighting Up the Pathways to Caspase Activation Using Bimolecular Fluorescence Complementation
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Lighting Up the Pathways to Caspase Activation Using Bimolecular Fluorescence Complementation

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使用CAS蛋白的RNA开关

Moe Hirosawa1, Hirohide Saito2

  • 1Department of Life Science Frontiers, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.

Methods in molecular biology (Clifton, N.J.)
|March 5, 2024
PubMed
概括

研究人员使用CRISPR-Cas系统设计了新的RNA结合蛋白 (RBPs). 这些Cas蛋白响应开关能够精确控制哺乳动物细胞中的基因翻译,扩展合成生物学工具.

科学领域:

  • 合成生物学 合成生物学
  • 分子生物学分子生物学
  • 基因调节 基因调节

背景情况:

  • 扩大RNA结合蛋白 (RBPs) 的库存对于开发哺乳动物细胞中复杂的转录后调节电路至关重要.
  • 克里斯普尔-卡斯系统提供了一个多功能平台,可以设计超越基因组编辑的新功能.
  • 目前用于转录后控制的合成生物学工具在范围和适用性方面存在限制.

研究的目的:

  • 使用CRISPR-Cas系统设计和验证新型RNA结合蛋白 (RBPs),用于转录后基因调节.
  • 为了证明Cas蛋白响应开关控制mRNA翻译的原理.
  • 扩大用于哺乳动物细胞合成生物学应用的工具包.

主要方法:

  • 通过将CRISPRRNA (crRNA) /单向导RNA (sgRNA) 序列纳入目标mRNA的5'未翻译区域 (5'UTR) 来设计的Cas蛋白响应基因开关.
  • 利用Cas蛋白的RNA结合能力,以Streptococcus pyogenes Cas9 (SpCas9) 为例,以调节翻译.
  • 开发了这些开关的基于等离子体和直接基于RNA的版本,以实现多样化的应用.
  • 在HEK293FT哺乳动物细胞系中验证了这些开关的功能.

主要成果:

  • 成功证明,Cas蛋白可以作为工程RBP来控制基因表达的功能.
关键词:
亚普特美尔 (Aptamer) 是一种药物.在CRISPRCas系统中.蛋白质响应开关 蛋白质响应开关在RBP中使用RBP.翻译法规的翻译法规

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  • 开发了功能性的Cas蛋白响应开关,可以调节mRNA翻译的调节.
  • 通过使用Streptococcus pyogenes Cas9 (SpCas9) 作为概念验证,展示了开关设计的适应性.
  • 在哺乳动物细胞环境 (HEK293FT细胞) 中证实了这些合成开关的有效性.

    • 结论:

    • 可以将CRISPR-Cas系统重新用于多功能RNA结合蛋白 (RBPs),以创建新的转录后调节工具.
    • Cas蛋白响应开关代表了合成生物学的一个强大而适应性的平台,可以精确控制基因翻译.
    • 这种方法显著扩大了可用的遗传部分,用于在哺乳动物系统中构建复杂的合成电路.