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相关概念视频

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

23
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...
23
CRISPR01:59

CRISPR

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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

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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...
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Homologous Recombination02:31

Homologous Recombination

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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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相关实验视频

Updated: Jul 13, 2025

Genome Editing in Mammalian Cell Lines using CRISPR-Cas
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Genome Editing in Mammalian Cell Lines using CRISPR-Cas

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在葡萄藤中使用CRISPR/LbCas12a系统进行高效的基因组编辑.

Chong Ren1,2,3, Elias Kirabi Gathunga1,2,3, Xue Li1,2,3

  • 1State Key Laboratory of Plant Diversity and Specialty Crops, Beijing Key Laboratory of Grape Sciences and Enology, Institute of Botany, the Chinese Academy of Sciences, Beijing, 100093, PR China.

Molecular horticulture
|October 18, 2023
PubMed
概括
此摘要是机器生成的。

现在,CRISPR/Cas12a系统,特别是LbCas12a,已经有效地用于葡萄藤基因组工程. 该系统成功诱导了目标基因的突变,热处理提高了效率,截断的crRNA显示出精确基因编辑的前景.

关键词:
克里斯普尔是什么意思?克里斯普尔是什么意思?黄类化合物 黄类在 LbCas12a 中.多重编辑多重编辑.葡萄酒 (Vitis vinifera) 是一种葡萄酒.长度crRNA长度crRNA长度crRNA长度crRNA长度crRNA长度

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CRISPR/Cas12a Multiplex Genome Editing of Saccharomyces cerevisiae and the Creation of Yeast Pixel Art
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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相关实验视频

Last Updated: Jul 13, 2025

Genome Editing in Mammalian Cell Lines using CRISPR-Cas
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CRISPR/Cas12a Multiplex Genome Editing of Saccharomyces cerevisiae and the Creation of Yeast Pixel Art
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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科学领域:

  • 植物生物技术 植物生物技术
  • 分子生物学分子生物学
  • 遗传学 遗传学 是一个

背景情况:

  • 集群定期间隔的短时间Palindromic重复 (CRISPR) /Cas12a系统在各种植物中提供了强大的基因组工程能力.
  • 葡萄藤 (Vitis vinifera) 尚未受益于CRISPR/Cas12a用于基因改造的应用.

研究的目的:

  • 开发和验证CRISPR/Cas12a系统,特别是Lachnospiraceae细菌ND2006 Cas12a (LbCas12a),用于葡萄藤细胞的向突变.
  • 调查基因淘汰对黄积累的影响,并通过环境和分子修改优化编辑效率.

主要方法:

  • 采用CRISPR/LbCas12a来准葡萄藤41B细胞中的塑性单糖转运体1 (TMT1) 和二黄-4-减少酶1 (DFR1) 基因.
  • 诱导了变异,并评估了热处理 (34°C) 对编辑效率的影响.
  • 分析了crRNA序列长度和截断crRNAs (trucrRNAs) 对基因组编辑结果的影响.

主要成果:

  • 通过敲除DFR1基因,在葡萄41B细胞中成功实现了向性突变发生,从而导致改变了黄类积累.
  • 热处理显著提高了TMT1的CRISPR/LbCas12a编辑效率,将其从35.3%提高到44.6%和从29.9%提高到37.3%.
  • 确定crRNA序列长度是编辑效率的关键因素,20nt trucrRNAs被证明与24nt crRNAs一样有效,而较短的序列 (≤18nt) 的效率较低.

结论:

  • CRISPR/LbCas12a系统是葡萄藤基因组工程的一个可行和有效的工具.
  • 优化热处理和crRNA设计等条件,包括使用 trucrRNAs,提高了葡萄藤基因编辑的精度和效率.
  • 这项研究为使用CRISPR/LbCas12a技术在葡萄藤中进行先进的基因操纵和作物改进铺平了道路.