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相关概念视频

CRISPR01:59

CRISPR

49.5K
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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CRISPR and crRNAs02:53

CRISPR and crRNAs

16.8K
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...
16.8K
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...
22.1K
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...
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Manipulation and Analysis01:21

Manipulation and Analysis

20
GIS manipulation and analysis functions are vital for decision-making and planning. These activities range from data retrieval tasks, such as selecting information based on specific criteria, to advanced analytical techniques that address complex spatial problems.One critical GIS analysis method is overlaying, which combines multiple data layers to examine impacts. For example, overlaying a river-dammed lake boundary with road networks can identify affected infrastructure. Another common...
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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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相关实验视频

Updated: Jun 7, 2025

Efficient PAM-Less Base Editing for Zebrafish Modeling of Human Genetic Disease with zSpRY-ABE8e
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工程CjCas9用于高效的基础编辑和主要编辑.

Siyuan Liu1, Yingdi Zhao1, Qiqin Mo1

  • 1Gene Editing Center, School of Life Science and Technology, ShanghaiTech University, Shanghai, China.

The CRISPR journal
|November 18, 2024
PubMed
概括
此摘要是机器生成的。

研究人员使用较小的Campylobacter jejuni Cas9蛋白开发了增强的CRISPR-Cas9基编辑器 (enCjBEs) 和主要编辑器 (enCjPEs). 这些新型编辑器显著提高了基因编辑效率,用于潜在的治疗应用.

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相关实验视频

Last Updated: Jun 7, 2025

Efficient PAM-Less Base Editing for Zebrafish Modeling of Human Genetic Disease with zSpRY-ABE8e
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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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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
  • 生物医学研究生物医学研究

背景情况:

  • 克里斯普尔-Cas9基因疗法面临着大Cas9蛋白大小的挑战,限制了腺相关病毒载体包装.
  • 链球菌 pyogenes Cas9 是常用的,但很难有效地提供.
  • 坎皮洛巴克特球菌Cas9 (CjCas9) 是一个较小的替代品,但其基编辑 (CjBE) 和主要编辑 (CjPE) 的效率有限.

研究的目的:

  • 从CjCas9中设计增强的CRISPR-Cas9基编辑器 (enCjBEs) 和主要编辑器 (enCjPEs),以改进基因编辑.
  • 提高CjCas9衍生基因编辑工具的效率和适用性.
  • 为生物研究和基因治疗开发紧而高效的编辑器.

主要方法:

  • 用P47K突变改造CjCas9以创建增强的细胞因子基编辑器 (enCjBEs) 和腺因基编辑器 (enCjABEs).
  • 使用CjCas9 P47K变种开发了增强的CjPE (enCjPE).
  • 融合了Sso7d和MS2的阿胺,产生了SsenCjPE,进一步优化了hMLH1dn和MMLV RTaseΔRnH,以及SsenCjPE-M2.2的D829R突变.

主要成果:

  • 实现了强大的C-to-T转换 (70%) 与enCjBEs和A-to-G转换 (76%) 与enCjABEs.
  • 与野生型CjPE相比,enCjPE在PRNP网站的编辑效率提高了17倍.
  • 在PRNP网站上,SsenCjPE实现了12%的平均编辑效率 (增加了24倍),而SsenCjPE-M2显示了61倍的效率.

结论:

  • 工程CjCas9变体 (enCjBEs,enCjABEs,enCjPEs,SsenCjPEs,SsenCjPE-M2) 提供了紧而高效的基因编辑解决方案.
  • 这些新的编辑器显著增强了基础和主要编辑功能.
  • 开发的工具对推进基因疗法和生物研究应用有前途.