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相关概念视频

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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...

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相关实验视频

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Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio
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一个高效的基于矢量CRISPR/Cas9系统在斑马鱼细胞系中.

Xiaokang Ye1, Jiali Lin2, Qiuji Chen1

  • 1Guangdong Provincial Key Laboratory of Marine Biotechnology, Institute of Marine Sciences, Shantou University, 243 Daxue Road, Shantou, 515063, China.

Marine biotechnology (New York, N.Y.)
|April 23, 2024
PubMed
概括
此摘要是机器生成的。

研究人员开发了一种新的CRISPR/Cas9基因编辑工具,用于斑马鱼细胞系. 这个系统有效地准鱼类细胞中的基因,使得高通量功能分析 in vitro.

关键词:
这就是CRISPR/Cas9的作用.鱼的细胞是鱼的细胞.在U6的推广者.斑马鱼是一种斑马鱼.

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科学领域:

  • 分子生物学分子生物学
  • 遗传学 遗传学 是一个
  • 生物技术是生物技术.

背景情况:

  • 克里斯普尔/卡斯9系统是一个强大的基因组编辑工具,但由于低效的促进器系统,它在鱼类细胞系中面临挑战.
  • 现有的载体通常需要单独的组件来表达sgRNA和Cas9,这阻碍了鱼类细胞的有效传递.

研究的目的:

  • 开发一个优化的全合一CRISPR/Cas9载体,用于在斑马鱼细胞系中高效的基因编辑.
  • 建立一个强大的体外平台,用于鱼类高通量基因功能分析.

主要方法:

  • 利用斑马鱼U6RNA聚合酶III (ZFU6) 促进体来驱动sgRNA和Cas9表达在单个lentiviral向量内.
  • 在lentiCRISPRV2系统的基础上构建了一个优化的向量,用ZFU6.6取代人类的U6促销器.
  • 测试了向量在斑马鱼纤维细胞 (PAC2) 细胞中向ctgfa基因以及向EMX1基因的哺乳动物293T细胞中的疗效.

主要成果:

  • 在斑马鱼胚胎中,ZFU6促进剂有效诱导了铁酶 (tyr) 基因编辑,导致视网膜色素的损失.
  • 优化的全合一载体成功准了PAC2细胞中的ctgfa基因,证明了其功能.
  • 该载体还在修改哺乳动物293T细胞中的EMX1基因方面表现出有效性,表明其广泛适用性.

结论:

  • 一个高效的CRISPR/Cas9基因编辑工具被建立为斑马鱼细胞系使用ZFU6促进器.
  • 这种新型的载体系统促进了鱼类细胞系中高通量基因功能分析.
  • 该工具显示了在包括哺乳动物细胞在内的不同细胞类型中广泛应用的潜力.