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相关概念视频

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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CRISPR01:59

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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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RNA Editing02:23

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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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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.
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: Jun 11, 2025

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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一个循环 permuted CasRx 平台,用于高效的,特定站点的RNA编辑.

Yuanming Wang1,2, Kaiwen Ivy Liu1,2, Mengying Mandy Liu1,2

  • 1School of Chemistry, Chemical Engineering and Biotechnology, Nanyang Technological University, Singapore, Singapore.

Nature biotechnology
|October 9, 2024
PubMed
概括

我们开发了xPERT,一个优化的RNA编辑平台,通过工程 CasRx. xPERT提供高在目标RNA编辑和低在目标之外的效果,使得精确的RNA序列修改没有基因组损伤.

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科学领域:

  • 分子生物学分子生物学
  • 生物技术是生物技术.
  • 在RNA治疗方面,RNA疗法.

背景情况:

  • 现有的RNA编辑工具面临着有效性和特异性的挑战.
  • 非目标RNA编辑仍然是开发可靠工具的重大障碍.

研究的目的:

  • 开发一个优化的RNA编辑平台,提高有效性和特异性.
  • 设计一个基于CasRx的工具,用于可编程RNA编辑.

主要方法:

  • 理性蛋白质工程的一个不活跃的Cas13正方体融合到一个突变的ADAR2除氨酶域.
  • 一个CasRx K940L突变体的循环 permutation,以创建一个强大的支架.
  • 与REPAIR系统对比开发的工具 (xPERT).

主要成果:

  • 设计的xPERT平台表现出强大的目标RNA编辑活动.
  • 与以前的系统相比,xPERT显著减少了目标之外的编辑.
  • 通过循环顺序的拓重新排列,为除氨酶域创造了一个强大的绑定支架.

结论:

  • xPERT平台为可编程RNA编辑提供了一个强大的解决方案.
  • 这项技术可以在没有基因组改变的情况下精确改变RNA序列.
  • xPERT可实现临时细胞修饰和定制的蛋白质功能.