利用高效的Exo:Cas12i3-5M融合,在大米中进行强大的单个和多重基因编辑
Wenxue Wang1,2, Shaoya Li1,2, Jiaying Yang1,2
1State Key Laboratory of Crop Gene Resources and Breeding, Institute of Crop Sciences (ICS), Chinese Academy of Agricultural Sciences (CAAS), Beijing, 100081, China.
Journal of integrative plant biology
|January 28, 2025
概括
研究人员通过将其与外核酶融合,增强了大米中的CRISPR/Cas12i3基因编辑. UL12:Cas12i3-5M融合显著提高了单个和多重编辑效率,为作物改进提供了强大的工具.
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
- 基因组编辑 基因组编辑
背景情况:
- 由于其小尺寸和灵活的PAM识别,CRISPR/Cas12i3是一个有前途的基因编辑系统.
- 以前的Cas12i3系统的编辑效率较低,限制了它们在植物多重基因编辑中的应用.
- 提高Cas12i3的效率对于功能基因组学和在作物改进中的金字塔化有益特征至关重要.
研究的目的:
- 利用CRISPR/Cas12i3.3开发一种更有效的单个和多重基因编辑系统,用于大米中.
- 系统地评估不同外核酶 (Exo) 融合与优化 Cas12i3 变体 (Cas12i3-5M) 的性能.
- 建立一个强大的基因编辑工具,通过金字塔化农业学重要特征来促进作物改进.
主要方法:
- 它将4个5'外核酶 (T5E,UL12,Pape,ME15) 融合到一个优化的Cas12i3变体 (Cas12i3-5M) 的N端.
- 系统地评估了这些Exo:Cas12i3-5M融合在米稳定线的六个内源性目标中的编辑效率.
- 评估了开发的融合系统的单个和多重基因编辑能力.
主要成果:
- 与Cas12i3.3-5M相比,Exo:Cas12i3-5M融合显著提高了基因编辑效率,高达12.46倍.
- UL12:Cas12i3-5M融合在六个目标中实现了高单基因编辑效率 (90.42%-98.61%).
- UL12:Cas12i3-5M表现出优异的多重编辑,三种基因的效率为82.76%,六种基因的效率为51.06%.
结论:
- UL12:Cas12i3-5M融合系统是一个强大的和高效的工具,用于单个和多重基因编辑在米中.
- 这种增强的系统扩展了现有的植物基因组编辑工具包.
- 它将有助于建立理想特征的金字塔,以加速作物改进.
更多相关视频
07:43Agrobacterium-Mediated Genetic Transformation, Transgenic Production, and Its Application for the Study of Male Reproductive Development in Rice
Published on: October 6, 2020
12.3K
09:51Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
Published on: May 25, 2018
33.7K
相关概念视频
02:31Homologous Recombination
50.1K
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...
50.1K
02:53CRISPR and crRNAs
16.6K
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...
16.6K