相关实验视频

Updated: Jul 3, 2025

CRISPR-Cas9-Mediated Genome Editing in the Filamentous Ascomycete Huntiella omanensis

CRISPR-Cas9-Mediated Genome Editing in the Filamentous Ascomycete Huntiella omanensis

Published on: June 9, 2020

在Eustoma grandiflorum中优化CRISPR-Cas9系统

Xueqi Li^1,2, Fanqi Bu^1,2, Lishan Wang^1,2

¹Key Laboratory of Saline-alkali Vegetation Ecology Restoration (Northeast Forestry University), Ministry of Education, Harbin 150040, China.

|February 16, 2024

概括

我们优化了Eustoma grandiflorum中的CRISPR-Cas9基因编辑系统,通过增强促销者和开发一个短暂的转换系统. 这种改进的系统实现了 EgPDS 基因的高编辑效率.

科学领域:

植物分子生物学植物分子生物学
基因编辑技术基因编辑技术
农业生物技术农业生物技术

背景情况:

优化CRISPR-Cas9系统对于提高植物基因编辑效率至关重要.
开发高效的转化系统对于Eustoma grandiflorum的遗传研究至关重要.
了解Eustoma grandiflorum中的基因功能需要强大的基因编辑工具.

研究的目的:

优化CRISPR-Cas9系统以提高Eustoma grandiflorum中的编辑效率.
为Eustoma grandiflorum建立一个高效的原生细胞分离和短暂转化系统.
应用修改后的CRISPR-Cas9系统来检测EgPDS基因的编辑效率.

主要方法:

在CRISPR-Cas9载体内优化单向导RNA和Cas9促进体.
为Eustoma grandiflorum建立一个高效的原生细胞分离和短暂转化协议.
修改CRISPR-Cas9系统的应用,以评估EgPDS基因编辑效率.

主要成果:

在Eustoma grandiflorum原生体中,EgU6-2促进体的活性大约是GmU6促进体的3倍.
经过修改的CRISPR-Cas9载体 (pEgU6-2::sgRNA-2/pEgUBQ10::Cas9-2) 实现了37.7%的编辑效率,比对照增加了30.3%.
由该系统诱导的突变包括基替代,小片段删除和插入.

结论:

成功开发了一种有效的基因编辑载体,用于Eustoma grandiflorum.
优化的CRISPR-Cas9系统为Eustoma grandiflorum基因功能研究提供了一个有价值的技术平台.
这项研究显著提高了Eustoma grandiflorum的基因编辑能力.

关键词:

分子生物学分子生物学植物生物学植物生物学植物生物技术植物生物技术

更多相关视频

Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution

Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution

Published on: February 26, 2019

Protocols for CRISPR/Cas9 Mutagenesis of the Oriental Fruit Fly Bactrocera dorsalis

Protocols for CRISPR/Cas9 Mutagenesis of the Oriental Fruit Fly Bactrocera dorsalis

Published on: September 28, 2022

相关概念视频

CRISPR

CRISPR

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...

CRISPR and crRNAs

CRISPR 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...