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相关概念视频

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

27
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...
27
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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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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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...
50.6K
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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相关实验视频

Updated: Jul 15, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

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通过PAM灵活的基因组编辑,使用工程化Cas9进行编辑.

Lin Zhao1, Sabrina R T Koseki1, Rachel A Silverstein2,3,4

  • 1Department of Biomedical Engineering, Duke University, Durham, NC, USA.

Nature communications
|October 4, 2023
PubMed
概括

研究人员通过结合SpRY和Sc+Cas9变体,设计出一种新的CRISPR酶Spryc. 这种嵌合体酶表现出高度灵活的原空间体相邻动机 (PAM) 识别,使得精确的基因组编辑跨多种序列的潜在治疗用途.

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相关实验视频

Last Updated: Jul 15, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

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科学领域:

  • 生物化学 生化学
  • 分子生物学分子生物学
  • 基因编辑技术的技术

背景情况:

  • 克里斯普尔酶需要一个特定的原空间器相邻动机 (PAM) 来识别目标部位.
  • 这一PAM要求限制了基因组编辑应用程序对某些DNA序列的可访问性.
  • 现有的Cas9变种在PAM灵活性,效率或准确性方面存在局限性.

研究的目的:

  • 为了设计一种具有增强和灵活的原空间器相邻动机 (PAM) 识别的仿真CRISPR酶.
  • 克服传统CRISPR-Cas9系统对序列可访问性的限制.
  • 开发一种多功能基因组编辑工具,用于各种治疗应用.

主要方法:

  • 重组SpRY的PAM交互域 (NRN>NYN PAM偏好) 与Sc++的N端 (NNG编辑功能).
  • 产生一种被指定为SpRYc的仿真酶,整合了两种母Cas9变体的特性.
  • 展示了SpRYc编辑多种PAM和与疾病相关的遗传位点的能力.

主要成果:

  • 仿真SpRYc酶表现出高度灵活和广泛的PAM识别能力.
  • SpRYc成功地和具体地编辑了各种原始空间器相邻图案 (PAM).
  • 该酶表明编辑与疾病相关的位置,表明潜在的治疗相关性.

结论:

  • 整合性蛋白质设计是设计先进的Cas9变体的强大策略.
  • 在PAM识别方面,SpRYc提供了强大的灵活性,扩大了CRISPR基因组编辑的范围.
  • 开发的酶激励下游应用程序,需要精确的基因组定位和编辑以前无法访问的网站.