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相关概念视频

CRISPR01:59

CRISPR

50.8K
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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CRISPR and crRNAs02:53

CRISPR and crRNAs

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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.
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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Homologous Recombination02:31

Homologous Recombination

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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...
50.5K
Epigenetic Regulation01:37

Epigenetic Regulation

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Epigenetic changes alter the physical structure of the DNA without changing the genetic sequence and often regulate whether genes are turned on or off. This regulation ensures that each cell produces only proteins necessary for its function. For example, proteins that promote bone growth are not produced in muscle cells. Epigenetic mechanisms play an essential role in healthy development. Conversely, precisely regulated epigenetic mechanisms are disrupted in diseases like cancer.
X-chromosome...
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Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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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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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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通过CRISPR/dCas9-Tet1介导的DNA甲基化编辑.

Junming Qian1, Shawn X Liu1

  • 1Department of Physiology and Cellular Biophysics, Columbia University Medical Center, Columbia University, New York, NY, USA.

Bio-protocol
|April 30, 2024
PubMed
概括

科学家们开发了一种针对性DNA甲基化编辑工具 (dCas9-Tet1),以精确地去甲基化特定的基因组位点. 这种系统可以研究表观遗传作用和微调基因表达,而不会改变DNA序列.

科学领域:

  • 表观遗传学 在表观遗传学中,表观遗传学是指表观遗传学.
  • 分子生物学分子生物学
  • 基因组学就是基因组学.

背景情况:

  • 基因甲基化是一种关键的表观遗传机制,涉及到生物过程和人类疾病.
  • 以前改变DNA甲基化的方法在很大程度上是非特异性的,阻碍了特定位置的功能研究.
  • 异常的DNA甲基化与各种人类疾病有关,需要精确的操纵工具.

研究的目的:

  • 开发一种用于在特定基因组位点进行向DNA脱甲基化的新工具.
  • 为了能够精确地调查DNA甲基化的功能意义.
  • 创建一个可扩展和高效的系统,用于细胞培养中的表观遗传操纵.

主要方法:

  • 十-十一转位二氧化酶1 (Tet1) 的催化域与一个失活的Cas9 (dCas9) 蛋白的融合.
  • 开发一个dCas9-Tet1系统用于由单导向RNA (sgRNA) 指导的序列特异性DNA脱甲基化.
  • 在各种细胞培养中建立一个模块化和可扩展的应用协议.

主要成果:

  • 在特定基因组位置的向DNA脱甲基化中表现出高效率和特异性.
  • 通过操纵DNA甲基化而使基因表达能够微调,而不会改变底层的DNA序列.
关键词:
克里斯普尔是什么意思?克里斯普尔是什么意思?通过DNA甲基化.在表观基因组编辑.基因调节 基因调节dCas9-Tet1 的使用情况

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  • 展示了dCas9-Tet1系统在各种细胞培养类型中的适用性.
  • 结论:

    • dCas9-Tet1系统为精确的表观遗传编辑提供了一个强大而通用的工具.
    • 这项技术有助于研究DNA甲基化在生物过程和疾病中的作用.
    • 该系统有可能在未来用于ex vivo和in vivo的治疗应用.