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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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Homologous Recombination02:31

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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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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
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相关实验视频

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Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines
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结合CRISPR激活和干扰能力使用dCas9和G-四重复结构.

Mohammad Lutful Kabir1, Sineth G Kodikara2, Mohammed Enamul Hoque1

  • 1Department of Chemistry and Biochemistry, Kent State University, Kent, OH 44242, USA.

bioRxiv : the preprint server for biology
|November 28, 2024
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概括
此摘要是机器生成的。

针对dCas9有效调节基因表达的c-Myc促进体中的G-四重复序列的CRISPR干扰和激活. 这种方法显示了控制c-Myc水平和细胞活力的潜力.

关键词:
克里斯普尔是什么意思?克里斯普尔是什么意思?克里斯普拉是一个.克里斯普里是什么?克里斯普里是什么?在G-quadruplex中使用.转录条例 转录条例 转录条例dCas9 的使用情况

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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:

  • 该c-Myc瘤基因在细胞增殖中起着至关重要的作用,在癌症中经常出现失调.
  • 发起地区的G-四重复形成序列 (PQS) 正成为关键的监管元素.
  • 基于CRISPR的工具提供了精确的基因组编辑功能.

研究的目的:

  • 为了研究利用核酶死Cas9 (dCas9) 对基因调节的c-Myc促进体向PQS的有效性.
  • 为了证明CRISPR干扰 (CRISPRi) 和CRISPR激活 (CRISPRa) 在c-Myc位置.
  • 阐明dCas9与PQS相互作用的机制细节及其对转录的影响.

主要方法:

  • 利用dCas9来准伯基特淋巴瘤细胞系中的c-Myc促进体中PQS附近的区域.
  • 通过针对非模板链来使用CRISPR干扰,通过针对模板链来激活CRISPR.
  • 进行了体外生物物理研究,以补充细胞测定和理解转录调制.

主要成果:

  • 用dCas9准模板链破坏了G-四重复的稳定性,使c-Myc mRNA和蛋白分别增加了2.1倍和1.6倍.
  • 用dCas9针对非模板链减少了c-Myc mRNA和蛋白质的1.8倍和2.5倍,双位点向实现了3.6倍和9.8倍的减少.
  • 细胞活力测试显示,非模板链向的细胞活力测试显示了相应的减少,表明了功能后果.

结论:

  • 通过CRISPR-dCas9对c-Myc促进体PQS的向,可以实现强大的基因激活和抑制.
  • 这项研究提供了一种新的策略,通过G-quadruplex操纵调节瘤基因表达.
  • 这些发现得到了体外生物物理数据的支持,为转录控制提供了机理性的见解.