相关实验视频

Updated: Jan 17, 2026

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Published on: June 16, 2017

应用多重CRISPR/Cas9策略来消除转基因植物中的选择标记物

Mohammed Rafi¹, Mohamed ElSiddig¹, Maitha Aldarmaki¹

¹Khalifa Center for Genetic Engineering and Biotechnology, Al Ain, United Arab Emirates.

Frontiers in genome editing

|September 19, 2025

概括

研究人员开发了一种CRISPR/Cas9方法,从转基因烟草中去除可选择标记基因 (SMG),创造无标记植物. 这一战略提高了转基因作物的生物安全性和接受度.

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
遗传学遗传学是一个

背景情况:

可选择标记基因 (SMG) 对于开发转基因植物至关重要.
然而,SMG引发了人们对生物安全,监管和公众看法的担忧.
需要有效的方法来消除SMG转型后.

研究的目的:

利用CRISPR/Cas9技术,精确地从转基因烟草中切除SMG.
评估SMG去除对植物发育和基因表达的效率和影响.
产生无标记物转基因植物,以提高商业化和接受度.

主要方法:

设计了一个带有四个导向RNA (gRNA) 的CRISPR/Cas9载体,以准SMG侧边区域.
含有DSRED SMG的转基因烟草叶片被用CRISPR载体重新转化.
通过光,PCR,测序和qPCR,分析再生芽的SMG损失.

主要成果:

大约10%的再生芽显示成功的SMG切除.
序列分析证实了SMG的删除,并确定了目标地点的小内.
没有SMG的植物表现出正常的生长,生育能力,并保持了感兴趣的基因的表达.
没有Cas9的,没有标记物的植物通过T1代的分离获得.

结论:

以CRISPR/Cas9为媒介的SMG切除是一种有效的策略,用于创建无标记物转基因植物.
这种方法不会对植物的发育或生育能力产生不利影响.
该方法为提高转基因作物的生物安全性和可销售性提供了可行的解决方案.

关键词:

这就是CRISPR/Cas9的作用.没有标记的转基因无标记物.在多基斯特龙tRNA-gRNA中.选择的基因标志物选择基因标志物烟草烟草的烟草烟草是什么

更多相关视频

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing

Published on: May 5, 2023

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

Published on: April 30, 2018

相关概念视频

CRISPR/Cas9 Genome Editing

CRISPR/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...

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...