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相关概念视频

Mismatch Repair01:20

Mismatch Repair

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Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
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Mismatch Repair01:36

Mismatch Repair

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Overview
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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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Proofreading01:31

Proofreading

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Synthesis of new DNA molecules is carried out by the enzyme DNA polymerase, which adds nucleotides on the daughter strand complementary to the template DNA strand. DNA polymerase has a higher affinity to add the correct base and ensures fidelity during DNA replication. Furthermore,  it exhibits proofreading activity during replication, using an exonuclease domain that cuts off incorrect nucleotides from the nascent DNA strand.
Errors During Replication are Corrected by the DNA Polymerase...
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Proofreading01:43

Proofreading

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Overview
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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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相关实验视频

Updated: Jan 19, 2026

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
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卷轴-卷轴异构体介导的分裂基编辑系统能够实现灵活和强大的核酸替代.

Shuangshuang Mu1,2,3, Qianru Li1,2,4,5, Menglong Chen6

  • 1China-New Zealand Joint Laboratory on Biomedicine and Health, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Institute of Development and Regeneration, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.

Nature communications
|January 17, 2026
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概括

研究人员开发了一种使用卷轴式卷轴异构体的分裂基编辑器 (BE) 系统,以克服腺相关病毒 (AAV) 包装的限制. 这种新的系统维持或提高了 in vivo 应用的基准编辑效率.

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A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
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相关实验视频

Last Updated: Jan 19, 2026

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A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
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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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科学领域:

  • 分子生物学分子生物学
  • 基因编辑技术的技术

背景情况:

  • 基础编辑器 (BEs) 提供精确的基础替代,但对于腺相关病毒 (AAV) 传递来说太大了.
  • 这种尺寸限制阻碍了它们在体内基因编辑疗法中的应用.

研究的目的:

  • 设计一个与AAV包装兼容的分割基编辑系统.
  • 评估新型分割基编辑系统的编辑效率和实体应用性.

主要方法:

  • 设计了一个分裂基编辑系统,通过通过卷轴-卷轴异构体将除氨酶与Cas9尼克酶融合.
  • 开发了基编辑器 (CC-CBE) 和基编辑器 (CC-ABE) 以及它们的衍生物.
  • 在各种细胞类型和体内小鼠模型中评估编辑效率.

主要成果:

  • 与未分割的BEs相比,分割的基础编辑器 (CC-BEs) 保持或提高了编辑效率.
  • 在初级体细胞中,CC-CBE实现了高达12.4倍的增强.
  • 在小鼠中,CC-ABE在使用双AAV载体的Pcsk9和Dmd基因中成功证明了体内A-to-G转换.

结论:

  • 开发了一个简单的,通用的分割基础编辑器策略,以克服AAV包装限制.
  • 该CC-BE系统使得有效的体内基编辑在不损害精度.
  • 这种方法扩大了基础编辑用于治疗应用的潜力.