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相关概念视频

CRISPR01:59

CRISPR

50.0K
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...
50.0K
RNA Editing02:23

RNA Editing

8.9K
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...
8.9K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.0K
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...
6.0K
CRISPR and crRNAs02:53

CRISPR and crRNAs

16.9K
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...
16.9K

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相关实验视频

Updated: Jun 16, 2025

CIRCLE-Seq for Interrogation of Off-Target Gene Editing
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CIRCLE-Seq for Interrogation of Off-Target Gene Editing

Published on: November 1, 2024

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设计了IscB-ωRNA系统,用于基础编辑扩大了目标范围.

Qingquan Xiao1,2, Guoling Li1, Dingyi Han2,3

  • 1HuidaGene Therapeutics Co. Ltd., Shanghai, China.

Nature chemical biology
|August 15, 2024
PubMed
概括

研究人员确定了新的IscB蛋白质用于基编辑,增强一个以扩大其准范围. 这些设计的IscB基编辑器对研究和治疗诸如杜申肌肉发育不良等遗传疾病有前途.

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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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Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors

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相关实验视频

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CIRCLE-Seq for Interrogation of Off-Target Gene Editing

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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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科学领域:

  • 分子生物学分子生物学
  • 基因编辑技术的技术
  • 微生物基因组学 微生物基因组学

背景情况:

  • IscB蛋白质是Cas9核酶的进化祖先,作为RNA引导的DNA内核酶和尼克酶起作用.
  • 它们的紧性使得它们适合用于基础编辑应用,但有限的准范围限制了它们的实用性.
  • 识别具有多种目标相邻动机 (TAM) 识别的IscB蛋白质对于扩展基编辑能力至关重要.

研究的目的:

  • 在哺乳动物细胞中发现具有活性的新型IscB蛋白.
  • 改造现有的 IscB 蛋白质以增强活性和扩大 TAM 识别.
  • 开发IscB衍生基编辑器用于治疗应用,包括遗传疾病纠正.

主要方法:

  • 从未培养的微生物中选非特征化的IscB蛋白质.
  • 蛋白质和 ωRNA 工程来增强 IscB 活动和改变 TAM 特异性.
  • 脱氨酶域与工程IscB尼克酶的融合,以创建基数编辑器.
  • 在哺乳动物细胞和疾病模型中的基编辑器效率的体外和体内验证.

主要成果:

  • 从19个未表征的微生物候选物中鉴定出10个活跃的IscB蛋白.
  • 设计了IscB.m16以创建IscB.m16*,将TAM范围从MRNRAA扩展到NNNGNA.
  • 开发了IscB.m16*衍生基编辑器,在哺乳动物细胞中表现出强大的效率.
  • 在小鼠中使用单个AAV输送的基编辑器成功恢复了杜恩肌肉衰竭蛋白.

结论:

  • 这项研究提供了一套来自新型和工程IscB蛋白质的紧基编辑工具.
  • 扩大了IscB.m16*的TAM范围,提高了其用于基因编辑的多功能性.
  • 基于IscB的基准编辑器对基因疾病的基础研究和治疗干预都有很大的潜力.