已发现的CRISPR-Cas12f同类驱动RNA引导的转录
Florian T Hoffmann1,2, Tanner Wiegand1,3, Adriana I Palmieri1
1Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY, USA.
Nature
|March 4, 2026
概括
研究人员发现了一种新的RNA引导基因激活机制,使用细菌Cas12f蛋白和sigma-E因子. 这种系统使得无需传统的促进子序列的可编程转录成为可能,为基因调节提供了新的可能性.
科学领域:
- 分子生物学分子生物学
- 基因规则 基因规则
- 细菌遗传学 细菌遗传学
背景情况:
- 细菌转录启动通常需要RNA聚合酶促进体识别的西格玛因子.
- 虽然大肠杆菌的SIGMA因子得到了很好的研究,但Bacteroidetes物种拥有众多未表征的SIGMA-E因子,这表明它们具有复杂的调节作用.
- 许多专业的西格玛-E因子的精确功能仍然是未知的.
研究的目的:
- 在细菌中发现RNA引导基因激活的新型机制.
- 研究西格玛-E因子与核酶死Cas12f (dCas12f) 的协调作用.
- 探索可编程的转录启动独立于正规的促进器动机.
主要方法:
- 在大肠杆菌中选dCas12f和sigma-E因子同类物.
- 使用RNA和染色体免疫沉实验来评估DNA标结合和指导RNA丰富.
- 同表达实验,以评估dCas12f-gRNA-sigma-E复合体对RNA聚合酶全酶的招募.
主要成果:
- 识别dCas12f-gRNA-sigma-E系统能够强大的DNA标结合,需要最小的5'-G动机.
- 证明dCas12f和指导RNA依赖于西格玛-E招募,表明直接的蛋白质-蛋白质相互作用.
- 三元复合物成功地驱动了强大的基因表达,没有促进体序列,转录起点由dCas12f介导的R循环决定.
结论:
- 已经发现了RNA引导转录启动的新范式.
- 这种机制利用dCas12f-gRNA-sigma-E复合体进行可编程的基因激活.
- 这些发现呈现出与CRISPR激活 (CRISPRa) 技术类似的自然RNA引导基因激活特征.
相关概念视频
CRISPR
58.4K
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...
58.4K
Homologous Recombination
64.6K
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
64.6K
CRISPR and crRNAs
19.4K
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...
19.4K
CRISPR/Cas9 Genome Editing
2.2K
The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
2.2K
RNA Interference
28.3K
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...
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...
28.3K


