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相关概念视频

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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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...
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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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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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RNA Editing02:23

RNA Editing

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RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
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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...
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Experimental RNAi02:15

Experimental RNAi

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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...
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相关实验视频

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A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
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一个分割的CRISPR/Cas13b系统用于条件RNA调节和编辑

Ying Xu1, Na Tian1, Huaxia Shi1

  • 1Department of Chemistry, Case Western Reserve University, 2080 Adelbert Road, Cleveland, Ohio 44106, United States.

Journal of the American Chemical Society
|February 22, 2023
PubMed
概括
此摘要是机器生成的。

研究人员开发了一种可通过酸和光控制的新CRISPR/Cas13b系统. 这种系统精确地调节RNA水平和修改,推进RNA研究工具.

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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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CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.
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CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

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相关实验视频

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科学领域:

  • 分子生物学
  • 生物技术
  • 核糖核酸生物学

背景情况:

  • CRISPR/Cas13b系统是RNA研究的一个强大工具.
  • 对Cas13b活动的精确控制对于理解RNA功能至关重要.
  • 现有的工具往往会干扰本地RNA活动.

研究的目的:

  • 为条件RNA调节设计一个分裂的Cas13b系统.
  • 为精确的RNA操纵开发一个ABA诱导系统.
  • 探索基于光的RNA修饰沉积的控制.

主要方法:

  • 通过酸 (ABA) 激活的分裂CRISPR/Cas13b系统的工程.
  • 开发一个可诱导 ABA 的 dCas13b 分离器,用于 m6A 沉积.
  • 使用可光激活的ABA衍生物进行光介导控制.
  • 证明了依赖于剂量和时间的RNA下调.

主要成果:

  • 使用ABA进行Cas13b/dCas13b活动的条件激活和禁用.
  • 在特定的RNA位点进行时间控制的m6A沉积.
  • 分离Cas13b/dCas13b系统活动的光诱导调制.
  • 在剂量和时间的依赖下调内源RNAs.

结论:

  • 开发的Cas13b/dCas13b分离系统提供了对RNA操纵的精确时间和条件控制.
  • 这些平台扩展了CRISPR和RNA调节工具包,
  • 这些系统最大限度地减少了内源RNA的功能破坏,促进了先进的RNA研究.