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相关概念视频

CRISPR01:59

CRISPR

50.5K
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...
50.5K
Regulation of Expression at Multiple Steps01:23

Regulation of Expression at Multiple Steps

889
The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the...
889
Experimental RNAi02:15

Experimental RNAi

6.1K
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...
6.1K
Regulation of Expression Occurs at Multiple Steps02:24

Regulation of Expression Occurs at Multiple Steps

22.6K
Gene expression can be regulated at almost every step from gene to protein. Transcription is the step that is most commonly regulated. This involves the binding of proteins to short regulatory sequences on the DNA. This association can either promote or inhibit the transcription of a gene associated with the respective sequence.
Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...
22.6K
RNA Interference01:23

RNA Interference

26.0K
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...
26.0K
CRISPR and crRNAs02:53

CRISPR and crRNAs

17.0K
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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相关实验视频

Updated: Jun 22, 2025

CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery
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CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery

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使用可编程,多输入处理CRISPR指导RNAs进行内源基因表达的逻辑调节.

Hansol Kang1, Dongwon Park1, Jongmin Kim1

  • 1Department of Life Sciences, Pohang University of Science and Technology, Pohang 37673, Korea.

Nucleic acids research
|June 29, 2024
PubMed
概括

工程条件导向RNAs (gRNAs) 提供了对大肠杆菌中的CRISPR-Cas系统的精确控制. 这一突破使得用于代谢工程和细胞形态学研究的动态基因调控成为可能.

科学领域:

  • 合成生物学 合成生物学
  • 分子生物学分子生物学
  • 微生物学 微生物学

背景情况:

  • 克里斯普尔-卡斯系统提供了灵活的RNA导向应用.
  • 工程指导RNAs (gRNAs) 允许对CRISPR-Cas.进行条件控制.
  • 对代谢过程的CRISPR-Cas精确调节是一个挑战.

研究的目的:

  • 开发一个强大的dCas9调节器与工程条件gRNAs.
  • 通过使用条件gRNAs来严格控制内源基因.
  • 在大肠杆菌中展示条件gRNA应用.

主要方法:

  • 工程条件gRNA与触发RNA相互作用,以控制基因表达.
  • 在大肠杆菌中使用了dCas9调节系统.
  • 评估基因表达动态范围和逻辑门操作 (A 或 (B 和 C)).
  • 向的内源代谢基因 (lacZ, malT, poxB) 和细胞骨基因 (ftsZ, mreB).

主要成果:

  • 有条件的gRNA在基因表达中达到高达130倍的动态范围.
  • 通过向代谢基因,证明了对代谢流和生长的控制.
  • 通过向细胞骨架基因来调节细胞丝和细胞分裂.

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A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
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A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization

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CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery
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Dual CRISPR-Interference Strategy for Targeting Synthetic Lethal Interactions Between Non-Coding RNAs in Cancer Cells
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A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
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A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization

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  • 实现了双输入逻辑门,用于条件 ftsZ 调节,诱导形态变化.
  • 结论:

    • 工程条件gRNAs提供精确的,可编程控制在大肠杆菌的基因表达.
    • 该系统能够调节细胞过程,包括新陈代谢和形态.
    • 这个平台可能与其他Cas-effector和宿主生物体兼容.