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相关概念视频

Long-patch Base Excision Repair01:02

Long-patch Base Excision Repair

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Since the discovery of the two BER pathways, there has been a debate about how a cell chooses one pathway over the other and the factors determining this selection. Numerous in vitro experiments have pointed out multiple determinants for the sub-pathway selection. These are:
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Base Excision Repair01:54

Base Excision Repair

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One of the common DNA damages is the chemical alteration of single bases by alkylation, oxidation, or deamination. The altered bases cause mispairing and strand breakage during replication. This type of damage causes minimal change to the DNA double helix structure and can be repaired by the base excision repair (BER) pathways. BER corrects damaged DNA sequences by removing the damaged base and restoring the original base sequence using the complementary strand as a template.
The first step of...
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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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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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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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基础编辑器的分割补充以最大限度地减少目标之外的编辑.

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概括

一个新的安全编辑 (SAFE) 系统的分裂除改进了基础编辑器 (BE),通过防止意外的DNA和RNA编辑. 这一突破提高了各种细胞类型的基因组编辑的精度和纯度.

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科学领域:

  • 分子生物学分子生物学
  • 基因组编辑 基因组编辑
  • 生物技术是生物技术.

背景情况:

  • 基因编辑器 (BEs) 对于在基因组中引入精确的点突变至关重要.
  • 传统的BEs在基因组和转录组中都表现出非目标编辑.
  • 现有的减轻脱效应的方法通常需要去氨酶突变或外部调节剂.

研究的目的:

  • 开发一种新的系统,以提高基础编辑的安全性和特异性.
  • 为了最大限度地减少指导RNA (gRNA) 独立和gRNA依赖的目标外编辑.
  • 提高BEs的整体目标编辑效率和产品纯度.

主要方法:

  • 为安全编辑 (SAFE) 系统开发一个分裂的除氨酶.
  • 在Cas9尼克酶的除氨酶域内分解BEs,以非激活两个组件.
  • 在植物,人类和酵母细胞中展示了gRNA受条件的重组,以进行有针对性的编辑.

主要成果:

  • 该SAFE系统有效地抑制了gRNA独立和gRNA依赖的非目标DNA和RNA编辑.
  • 在各种细胞类型 (植物,人类,酵母) 中实现了强大的目标编辑.
  • 显著提高产品的纯度,由于消除了indels.

结论:

  • 在基础编辑器中,SAFE系统提供了一个可通用的策略来减少非目标编辑.
  • 这种方法提高了基因组编辑应用程序的精度和可靠性.
  • SAFE为更安全,更有效的基础编辑提供了强大的解决方案.