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相关概念视频

CRISPR01:59

CRISPR

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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...
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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...
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CRISPR and crRNAs02:53

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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.
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Cooperative Binding of Transcription Regulators02:13

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Transcriptional regulators bind to specific cis-regulatory sequences in the DNA to regulate gene transcription. These cis-regulatory sequences are very short, usually less than ten nucleotide pairs in length. The short length means that there is a high probability of the exact same sequence randomly occurring throughout the genome.  Since regulators can also bind to groups of similar sequences, this further increases the chances of random binding. Transcriptional regulators form...
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Gene transcription is regulated by the synergistic action of several proteins that form a complex at a gene regulatory site. This is observed in eukaryotes, where the regulation of gene expression is a complex process. Regulatory proteins in eukaryotes can broadly be classified into two types – regulators that bind directly to specific DNA sequences and co-regulators that associate with regulatory proteins but cannot directly bind to the DNA. These co-regulators are further divided into...
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Transcription activators are proteins that promote the transcription of genes from DNA to RNA. In most cases, these proteins contain two separate domains ‒ a domain that binds to DNA and a domain for activating transcription; however, in some cases, a single domain is responsible for both binding and activation of transcription, as seen in the glucocorticoid receptor and MyoD.
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相关实验视频

Updated: Jun 15, 2025

Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines
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特定的多价值分子可以促进CRISPR介导的转录激活.

Rui Chen1,2,3,4, Xinyao Shi1,3, Xiangrui Yao1,3

  • 1Shenzhen Key Laboratory of Gene Regulation and Systems Biology, School of Life Sciences, Southern University of Science and Technology, Shenzhen, China.

Nature communications
|August 22, 2024
PubMed
概括

内在无序区域 (IDR) 和模块化域 (MD) 可以增强CRISPR/Cas基因激活. 最佳的合作性,而不是最大的合作性,以及针对促进剂-增强剂相互作用是使用这些多价值分子进行强大的激活的关键.

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科学领域:

  • 分子生物学分子生物学
  • 基因规则 基因规则
  • 生物技术是生物技术.

背景情况:

  • 克里斯普尔/卡斯系统提供精确的基因编辑能力.
  • 与CRISPR/Cas组件融合的转录激活剂可以调节基因表达.
  • 本质上无序区域 (IDR) 和模块化域 (MD) 在增强CRISPR/Cas活动中的作用尚未完全被理解.

研究的目的:

  • 研究IDRs增强CRISPR/Cas基转录激活的机制.
  • 探索将IDRs和MDs与CRISPR/Cas激活剂相结合的协同效应.
  • 通过使用工程CRISPR/Cas系统,确定强大的基因激活的最佳策略.

主要方法:

  • 将12种不同的IDR与dCas9-VP64转录激活剂融合.
  • 评估IDRs对基因激活的影响,独立于相位分离.
  • 将dCas9-VP64-IDR结构与模块化域 (MD) 结合起来.
  • 系统地改变gRNA结合位点并评估cis-trans合作性.
  • 准促进剂-增强剂对并评估染色质相互作用.

主要成果:

  • 在12种测试的IDR中,有7种增强了dCas9-VP64的激活,而不管分相.
  • 单独使用MDs并没有增强激活,但与dCas9-VP64-IDR结合时显著增强了激活.
  • 最佳的cis-trans合作性,而不是最大的,导致了最强大的基因激活.
  • 准促进剂-增强剂区域和增强色素相互作用放大了协同效应.

结论:

  • 通过结合IDRs和MDs,可以有效地增强工程CRISPR/Cas激活器.
  • 该研究提供了对多价值分子介导的基因激活机制的见解.
  • 使用CRISPR/Cas技术开发了一种用于有效激活基因的多功能平台.