在Fraxinus mandshurica的基因组编辑中开发一种内生促销器驱动的CRISPR/Cas9系统
Shangzhu Gao1,2, Mengfan Zhao1,2, Siyu Sun1,2
1State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040, China.
Forestry research
|September 19, 2025
概括
这项研究开发了一种高效的CRISPR/Cas9基因组编辑系统,用于满洲 (Fraxinus mandshurica) 树. 优化的促进剂,sgRNA和热处理显著提高了树木改善和林业可持续性的编辑效率.
科学领域:
- 植物生物技术 植物生物技术
- 基因组编辑 基因组编辑
- 林业科学 林业科学
背景情况:
- 克里斯普尔/Cas9技术彻底改变了树木改进,使得精确的特征修改和可持续林业成为可能.
- Fraxinus mandshurica是一种有价值的树种,对中国的林业计划至关重要,但需要高效的基因组编辑工具.
- 开发特定物种的基因组编辑系统对于像F. mandshurica.这样的反抗性物种至关重要.
研究的目的:
- 为Fraxinus mandshurica.开发和优化一个特定物种的CRISPR/Cas9基因组编辑平台.
- 通过内源促进体工程,sgRNA优化和受控的环境条件来提高基因组编辑效率.
- 建立一个功能性基因组学和F. mandshurica. 的特征改善的强有力的系统.
主要方法:
- 为了增强sgRNA表达,设计了截断的内源性FmU6促进体变体 (FmU6-6-4,FmU6-7-4).
- 利用内源性FmECP3促进剂用于高活性Cas9表达.
- 在转化和再生过程中,优化了针对FmPDS1/2的sgRNA,采用了热处理 (37°C),并调节了光质.
主要成果:
- FmU6促进子变体增加了sgRNA表达的3.11-3.36倍,比AtU6-26.
- 对于FmECP3促进体,其活性是Cas9表达的阳性对照的5.48倍.
- 在优化条件下,FmPDS1/2的裂解效率达到36.10%,热处理增加了Cas9活性7.77倍.
- 产生了具有18.2%编辑效率的仿真白色突变体,表现出减少的叶绿素含量 (46.44%-58.88%).
结论:
- 一个优化的CRISPR/Cas9系统显著提高了Fraxinus mandshurica的基因组编辑效率.
- 开发的平台有助于进行功能基因组学研究和改善这种有价值的树木物种的特征.
- 这项研究为推进林业生物技术和可持续森林管理提供了一个强大的工具.
相关概念视频
CRISPR/Cas9 Genome Editing
1.7K
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...
1.7K
CRISPR
57.5K
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...
57.5K
CRISPR and crRNAs
18.7K
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...
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...
18.7K
Homologous Recombination
62.7K
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...
62.7K


