在特定基因突入和细菌植物酶基因表达中的特定基因突入和细菌植物酶基因表达
Hassan Zadabbas Shahabadi1,2,3, Arash Akbarzadeh1, Hamideh Ofoghi2
1Department of Fisheries, Faculty of Marine Science and Technology, University of Hormozgan, Bandar Abbas, Iran.
Frontiers in plant science
|June 5, 2023
概括
通过CRISPR-Cas9基因编辑,成功地将一种细菌植物酶基因插入了Chlamydomonas reinhardtiitii.
科学领域:
- 分子生物学分子生物学
- 基因工程是一种基因工程.
- 生物技术是生物技术.
背景情况:
- 由于插入部位效应和基因沉默,基因插入可能具有挑战性.
- 在Chlamydomonas reinhardtii中外来基因的稳定表达对于商业应用至关重要.
研究的目的:
- 应用CRISPR-Cas9介导的同质导向DNA修复 (HDR) 进行精确的基因敲定.
- 将细菌植物酶基因插入克拉米多马纳斯 (Chlamydomonas reinhardtii) 中的酸盐减少酶 (NR) 基因.
- 为了确保转基因的稳定性,避免基因沉默.
主要方法:
- 使用CRISPR-Cas9核糖蛋白 (RNP) 复合体进行直接输送.
- 针对NR基因 (外显子2) 进行植物酶基因盒的特定插入.
- 通过PCR,测序和RT-qPCR确认编辑.
主要成果:
- 在24个Chlamydomonas reinhardtii克隆中成功将细菌植物酶基因敲入NR基因.
- 编辑效率为14.81%,基因磁带精确地位于exon 2.
- 经过10代的稳定转基因表达,没有基因沉默或负插入效应.
结论:
- 克里斯普尔-卡斯9介导的敲进是一种有效的工具,用于切莱米多蒙纳斯 Reinhardtii的特定部位的核基因表达.
- 这种方法避免了位置效应和基因沉默,确保了转基因的稳定性.
- RNP-CRISPR/Cas9基因编辑为加速C. reinhardtii的重组蛋白质生产提供了一个有希望的方法.
相关概念视频
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
Transgenic Plants
7.3K
Recombinant DNA technology called transgenesis is often used to add a foreign gene or remove a detrimental gene from an organism. Such genetically modified organisms are called transgenic organisms.
The first-ever transgenic plant was a tobacco plant developed in 1983 that showed resistance against the tobacco mosaic virus. Since then, many transgenic plants have been developed and commercialized for improving the agricultural, ornamental, and horticultural value of a crop plant. Transgenic...
The first-ever transgenic plant was a tobacco plant developed in 1983 that showed resistance against the tobacco mosaic virus. Since then, many transgenic plants have been developed and commercialized for improving the agricultural, ornamental, and horticultural value of a crop plant. Transgenic...
7.3K
Reporter Genes
11.6K
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
11.6K
Transgenic Organisms
31.4K
Overview
31.4K
In-vitro Mutagenesis
14.0K
To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
14.0K


