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相关概念视频

Long-patch Base Excision Repair01:02

Long-patch Base Excision Repair

7.0K
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:
7.0K
Proofreading01:31

Proofreading

6.3K
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...
6.3K
Mismatch Repair01:20

Mismatch Repair

4.8K
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...
4.8K
Base Excision Repair01:54

Base Excision Repair

22.3K
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...
22.3K

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相关实验视频

Updated: Jun 25, 2025

Design and Use of Multiplexed Chemostat Arrays
19:40

Design and Use of Multiplexed Chemostat Arrays

Published on: February 23, 2013

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开发多重合直角基编辑器 (MOBE) 系统.

Quinn T Cowan1, Sifeng Gu1, Wanjun Gu2

  • 1Department of Chemistry and Biochemistry, University of California San Diego, La Jolla, CA, USA.

Nature biotechnology
|May 21, 2024
PubMed
概括

新的基编辑器 (BEs) 允许精确,同时编辑DNA,而无需双链断裂. 这一突破克服了以前的局限性,使多重基因编辑用于研究和疾病建模.

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Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors
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Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors

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BEST: Barcode Enabled Sequencing of Tetrads
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BEST: Barcode Enabled Sequencing of Tetrads

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相关实验视频

Last Updated: Jun 25, 2025

Design and Use of Multiplexed Chemostat Arrays
19:40

Design and Use of Multiplexed Chemostat Arrays

Published on: February 23, 2013

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Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors
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Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors

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BEST: Barcode Enabled Sequencing of Tetrads
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BEST: Barcode Enabled Sequencing of Tetrads

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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
  • 生物技术是生物技术.

背景情况:

  • 基编辑器 (BEs) 提供精确的点突变安装,没有DNA双链断裂.
  • 由于指导RNA交叉和非对角编辑,多重化不同的BE类型 (腺因和细胞因) 是具有挑战性的.
  • 目前的方法限制了多个基准编辑器同时应用到不同的位置.

研究的目的:

  • 为多重应用设计直角腺因和细胞因基编辑器.
  • 为了克服指导RNA交叉的限制,同时编辑基础.
  • 为了在同一DNA链上的多个位置进行精确的同时发生的编辑.

主要方法:

  • 通过使用RNA胺蛋白-涂层蛋白系统进行酶招募的氨酸和氨酸基编辑器.
  • 开发了四个多重合并的正角形基数编辑系统.
  • 使用光丰富策略来提高同时发生的编辑速率.
  • 测试了系统兼容性与扩展的protospacer相邻动图和高保真Cas9变体.

主要成果:

  • 在相同的DNA链中实现了高达7.1%的精确并发编辑,而无需在同一DNA链中进行丰富.
  • 光丰富使人类细胞的同时发生的编辑率增加到24.8%.
  • 与各种Cas9变体和多种细胞类型的疗效证明了兼容性.
  • 与家长基准编辑系统相比,观察到同等或减少的目标外效应.

结论:

  • 成功开发了多重合并的直角基数编辑器,克服了以前的交叉语音限制.
  • 这些工程系统允许精确,同时安装多点突变.
  • 该技术有助于对疾病相关的点突变组合进行建模,提高了效率和安全性.