在瓜子中通过毛的根转化系统验证CRISPR构造活性和基因功能
Xiang Li1,2,3, Chenchen Cao2, Pablo Bolaños-Villegas4,5,6
1College of Landscape Architecture and Art, Henan Agricultural University, 63 Nongye Road, Zhengzhou, 450002 China.
概括
一种新方法使用瓜子 (Cucumis melo L.) 中的工程毛发根来快速测试基因编辑工具并分析基因功能,加速对这一重要的水果作物的研究.
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
- 农业科学 农业科学
背景情况:
- 瓜子 (Cucumis melo L.) 是一种全球种植的水果作物,具有重要的营养价值.
- 有效的基因编辑协议对于加速瓜子的功能基因分析至关重要.
- 毛发根诱导为植物遗传研究提供了一个快速的系统.
研究的目的:
- 开发一种有效的协议,用于诱导和转化瓜子中的毛发根.
- 建立一个快速验证CRISPR/Cas9基因编辑结构在植物中的系统.
- 分析瓜子基因,如CmRHL1在根部发育中的功能.
主要方法:
- 瓜子花皮被用作诱导毛发根的扩展剂,使用Agrobacterium rhizogenes K599.
- 为了评估基因编辑效率,CRISPR/Cas9构造被转化为诱导的毛发根.
- 评估了sgRNA表达卡塞特号对基因编辑效率的影响.
- 通过基因编辑来研究CmRHL1在根毛发育中的功能.
主要成果:
- 在一个月内,毛发根诱导 (68.61%) 和转换的高效率得到了实现.
- 该系统允许在植物中快速评估CRISPR/Cas9目标部位活动.
- 基因编辑效率与sgRNA表达盒的数量呈正相关.
- CmRHL1的突变显著抑制了西瓜根毛发的发育.
结论:
- 已建立的毛根编辑方法为快速的CRISPR/Cas9构造验证提供了一个补充工具.
- 这个系统在根部发育过程中促进了有效的基因功能表征.
- 该协议加速了瓜子的功能基因组学研究,补充了遗传基因组编辑方法.
相关概念视频
01:59CRISPR
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 Short...
02:53CRISPR and crRNAs
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...
01:59CRISPR
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 Short...
01:23The Antiviral System of Bacteria and Archaea: CRISPR
CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this defense.
01:28CRISPR/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...