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相关概念视频

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

39
The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
39
CRISPR01:59

CRISPR

52.1K
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...
52.1K

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相关实验视频

Updated: Jul 15, 2025

A Universal Protocol for Large-scale gRNA Library Production from any DNA Source
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A Universal Protocol for Large-scale gRNA Library Production from any DNA Source

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闪光基因组编辑管道:一种高效和高通量方法,用于构建数组化的CRISPR库,用于植物功能基因组学.

Lu Yao1, Xiaochun Wang1,2, Runnan Ke1

  • 1National Key Laboratory of Crop Genetic Improvement and Hubei Hongshan Laboratory, Huazhong Agricultural University, Wuhan, People's Republic of China.

Current protocols
|September 27, 2023
PubMed
概括

我们开发了FLASH基因组编辑管道,用于在植物中进行高通量CRISPR选. 这种方法使用独特的FLASH标签将基因目标与Cas9/gRNA载体联系起来,从而实现高效的基因功能发现.

关键词:
这就是CRISPR/Cas9的作用.排列式图书馆 排列式图书馆基因组编辑 基因组编辑高吞吐量,具有高吞吐量.植物植物植物植物植物.

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相关实验视频

Last Updated: Jul 15, 2025

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科学领域:

  • 植物基因组学 植物基因组学
  • 基因组编辑技术的技术
  • 农作物育种 农作物育种

背景情况:

  • CRISPR/Cas9是各种生物体中基因编辑的强大工具.
  • 使用CRISPR/Cas9的高通量基因查在植物中具有挑战性.
  • 需要有效的方法来进行植物功能基因组学和作物改进.

研究的目的:

  • 建立一个新的管道,用于在植物中构建阵列的CRISPR库.
  • 为了实现高通量遗传查和在植物中发现基因功能.
  • 开发一种方法,使用独特标识符将目标基因与Cas9/gRNA载体联系起来.

主要方法:

  • 开发FLASH基因组编辑管道,使用12个不同的PCR片段 (FLASH标签) 来索引Cas9/gRNA载体.
  • 组装FLASH标记索引的Cas9/gRNA等离子体,并准备一个等离子体库.
  • 米植物的农细菌媒介转化,使用索引菌株的混合物.
  • 使用常规PCR和凝电泳来分配基因编辑植物中的gRNA信息.

主要成果:

  • 在植物中使用FLASH基因组编辑管道成功构建了一个数组化的CRISPR库.
  • 建立了目标基因/gRNA和FLASH标签之间的独特联系,以有效地检索信息.
  • 证明了在细菌菌株和基因编辑植物中读取gRNA信息的能力.
  • 该协议允许在植物中快速和全面地发现基因功能.

结论:

  • FLASH基因组编辑管道促进了植物的高通量遗传查.
  • 这种方法可以有效地发现基因功能,并支持作物育种的进步.
  • 该方法为在植物系统中构建和分析数组CRISPR库提供了强大的解决方案.