エシェリキア・コライのゲノムスケールメタボリックリワイヤリングのための二次モードCRISPRa/i
Soo Young Moon1,2, Mi Ri Kim3, Nan-Yeong An1
1Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Republic of Korea.
Nucleic acids research
|August 22, 2025
まとめ
研究者らは,細菌の精密な遺伝子制御のために,二重モードのCRISPR活性化および干渉 (CRISPRa/i) システムを開発した. この新しいシステムは 細菌の代謝工学を進めて 同時に遺伝子を活性化し抑制します
科学分野:
- 合成生物学
- 分子生物学
- 微生物工学
背景:
- CRISPRによる転写制御は プログラム可能な遺伝子制御を提供します
- バクテリアの同時遺伝子活性化と抑制は,限られた活性化ドメインのために困難です.
研究 の 目的:
- バクテリアの精密な遺伝子調節のための効率的な二重モードのCRISPR活性化および干渉 (CRISPRa/i) システムを開発する.
- 遺伝子の活性化と抑制を 同時に行うための 汎用的なエフェクターを設計する
主な方法:
- PAMフレキシブルなdxCas9をエシェリキア・コリのcAMP受容体タンパク質 (CRP) と統合した.
- エフェクタ機能の強化のために,体系的に最適化されたCRPドメインとリンク.
- 検証と適用のために二重光レポーターとプールされたガイドRNAライブラリを使用した.
主要な成果:
- 精密な遺伝子活性化と抑制のための強力なdxCas9-CRPシステムを開発しました.
- 二重光レポーターを使用した複数の遺伝子の同時調節が実証された.
- ゲノムスケールのCRISPRa/iスクリーニングを通じて,重要な規制標的を特定することで,E. coliにおけるヴィオラセインの生産を成功させました.
結論:
- 開発された二重モードのCRISPRa/iシステムは,細菌の遺伝子調節を精密に柔軟に制御します.
- このシステムは 細菌の代謝経路の再配線を バイオテクノロジーの用途として進めている
- ゲノムスケールの調整された活性化と抑制を可能にします.
関連する概念動画
Stringent Response in E. coli
52
Bacterial growth is closely tied to nutrient availability, with cells proliferating exponentially under favorable conditions and entering a stationary phase when resources become scarce. This transition is mediated by a regulatory mechanism known as the stringent response, which allows bacteria to adapt to nutrient deprivation by modulating gene expression and metabolic activity.During nutrient scarcity, intracellular amino acid levels decline. It results in the accumulation of uncharged tRNAs...
52
Inducible Operons: lac Operon
141
The lac operon in Escherichia coli is a model for understanding inducible gene regulation and metabolic flexibility. It integrates local control by lactose and global regulation through catabolite repression, enabling E. coli to preferentially metabolize glucose when available and switch to lactose utilization when glucose is scarce.Structure and Function of the lac OperonThe lac operon contains three structural genes: lacZ (β-galactosidase), lacY (lactose permease), and lacA...
141
Other Glycolytic Pathways
214
The pentose phosphate pathway (PPP) operates in parallel with glycolysis, facilitating the metabolism of both pentoses and glucose. This pathway consists of two distinct phases: the oxidative and non-oxidative phases. While it does not directly generate ATP, the intermediates formed during the process can integrate into glycolysis, contributing to cellular energy metabolism when required.Oxidative Phase: NADPH ProductionThe oxidative phase of the pentose phosphate pathway is primarily...
214
CRISPR and crRNAs
17.4K
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...
17.4K
CRISPR
52.9K
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...
52.9K
Coordination of Gene Expression Processes in Bacteria
148
The DNA replication, transcription, and translation processes are intricately coupled in bacteria, allowing efficient gene expression and rapid protein synthesis. While this physical and functional coordination is advantageous, it introduces challenges that bacteria overcome through specific regulatory mechanisms.Coupling of Replication, Transcription, and TranslationThe coupling of replication, transcription, and translation is a hallmark of bacterial gene expression. As the replisome unwinds...
148


