関連する実験動画

Updated: Feb 19, 2026

Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution

Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution

Published on: January 16, 2017

標的RNA結合チャネルを覆うCas10の残基は,相性標的RNAと不一致の標的を区別することによって干渉を調節する

Sarah Khweis¹, Mason Blackburn¹, Calvin Perdigao¹

¹Department of Chemistry and Biochemistry, University of Alabama, Tuscaloosa, AL, USA.

|February 18, 2026

まとめ

タイプIIIのCRISPRシステムは,Cas10タンパク質を使用して,外来RNAを検出します. Cas10の特定の残基は干渉を調節し,防御を強化するために正しいと間違った標的RNAの差別を改善します.

科学分野:

分子生物学は分子生物学である.
微生物学微生物学とは
バイオケミストリーバイオケミストリー

背景:

Cas10タンパク質によって特徴づけられるCRISPRシステムIII型は,自然界で広く存在しています.
Cas10は外来RNAを細胞に検知する複合体を形成し,検出時に干渉カスケードを開始します.
Cas10による循環性オリゴアデニラート (cOA) 合成は,この干渉プロセスにとって極めて重要です.

研究の目的:

Cas10媒介の標的RNAセンシングの分子メカニズムを解明する.
Cas10における重要な残基を特定し,同種の標的RNAと不一致の標的RNAを区別する.
標的RNA結合ライセンスのCas10干渉活動の仕組みを理解する.

主な方法:

*S. epidermidis*からのCas10標的RNA結合チャネルにおける5つの重要な残留物の特定.
これらの残基が,同類および不一致の標的RNAとの干渉に与える寄与のインビヴォ評価.
局所指向のCas10-Csm変異体を用いたインビトロcOA合成アッセイ.

主要な成果:

特定された5つの残基は,標的RNAの互補性に基づいて干渉活性化を調節する.
これらの残留物の突然変異は,同族および不一致のcrRNA-標的RNA複合体間の差別を高めます.
インビトロアッセイでは,Cas10変異体がcOA合成における特異性の向上を示していることが確認されました.

結論:

Cas10の標的RNA結合チャネル内の特定の残基は,正確な標的認識に不可欠です.
これらの残留物は,干渉の活性化を調節し,オフターゲット効果を防止します.
この発見は,CRISPR型IIIシステムの特異性とエンジニアリングの可能性に関するメカニズム的な洞察を提供します.

キーワード:

CRISPR-Casは,CRISPR-Casをベースに開発されている.カス10 カス10 周期性オリゴアデニラート干渉干渉は干渉干渉である.タイプIIIはタイプIIIです.

さらに関連する動画

Exploring Sequence Space to Identify Binding Sites for Regulatory RNA-Binding Proteins

Exploring Sequence Space to Identify Binding Sites for Regulatory RNA-Binding Proteins

Published on: August 9, 2019

Author Spotlight: A Computational Pipeline for Analyzing Chimeric Noncoding RNA-Target RNA Interactions in High-Throughput Sequencing Data

Author Spotlight: A Computational Pipeline for Analyzing Chimeric Noncoding RNA-Target RNA Interactions in High-Throughput Sequencing Data

Published on: December 1, 2023

関連する概念動画

RNA Interference

RNA Interference

RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...

Riboswitches

Riboswitches

Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...

Types of RNA

Types of RNA

Overview
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...

Types of RNA

Types of RNA

Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in regulating gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA Performs Diverse...

Nucleic Acid Structure

Nucleic Acid Structure

The pentose sugar in DNA is deoxyribose, while in RNA the pentose sugar is ribose. The difference between the sugars is the presence of the hydroxyl group on the ribose's second carbon and a hydrogen on the deoxyribose's second carbon. The phosphate residue attaches to the hydroxyl group of the 5′ carbon of one sugar and the hydroxyl group of the 3′ carbon of the sugar of the next nucleotide, which forms a 5′ to 3′ phosphodiester linkage.
DNA Structure
DNA...

Experimental RNAi

Experimental RNAi

RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...