如何在植物中使用CRISPR/Cas9:从目标部位选择到DNA修复
Adéla Přibylová1, Lukáš Fischer1
1Department of Experimental Plant Biology, Faculty of Science, Charles University, Viničná 5, 12800, Prague 2, Czech Republic.
Journal of experimental botany
|April 22, 2024
概括
克里斯普尔/卡斯9基因组编辑提供了精度,但也有局限性. 了解影响Cas9活动和DNA修复的因素对于各种生物体的成功基因编辑结果至关重要.
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:
- 自2012年以来,集群定期间隔的短平行列重复 (CRISPR) /CRISPR相关蛋白 (Cas) 系统,特别是CRISPR/Cas9,已经彻底改变了基因组编辑.
- 尽管CRISPR/Cas9技术具有巨大的实力,但它面临着一些局限性,这些局限性影响了其在研究和应用环境中的效率和特异性.
研究的目的:
- 本次审查重点关注的是Streptococcus pyogenes的CRISPR/Cas9变种.
- 它旨在阐明影响基因组编辑结果的关键因素.
- 为优化植物基因组编辑提供了指导方针.
主要方法:
- 该审查分析了影响CRISPR/Cas9活性的因素,包括目标序列,染色质状态和Cas9变体持久性.
- 它检查了影响DNA修复机制的因素,例如细胞类型,细胞循环阶段和DNA末端类型 (形/分阶段).
- 在植物,酵母和动物中比较CRISPR/Cas9应用.
主要成果:
- 克里斯普尔/Cas9活动是由目标序列特征,染色质可访问性和Cas9存在的持续时间调节的.
- 受细胞类型和细胞周期影响的DNA修复途径选择,显著影响突变发生的结果.
- 对于CRISPR/Cas9应用的跨国知识传输是有限的,需要对特定生物体进行考虑.
结论:
- 对影响CRISPR/Cas9活动和DNA修复的因素的认识提高了基因组编辑的可预测性.
- 优化CRISPR/Cas9策略需要考虑特定生物体的生物背景.
- 提供了详细的指南,以提高植物基因组编辑的成功率.
相关概念视频
CRISPR
50.8K
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...
50.8K
Homologous Recombination
50.5K
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...
50.5K
CRISPR and crRNAs
17.0K
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...
17.0K
Conservative Site-specific Recombination and Phase Variation
6.0K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
6.0K


