相关实验视频

Updated: Sep 11, 2025

Genome Editing with CompoZr Custom Zinc Finger Nucleases ZFNs

Genome Editing with CompoZr Custom Zinc Finger Nucleases ZFNs

Published on: June 14, 2012

使用两个ZFN单体的T2A合共表达的高效基因组编辑

Shota Katayama¹, Takashi Yamamoto^1,2

¹Genome Editing Innovation Center, Hiroshima University, Higashi-Hiroshima 739-0046, Japan.

International journal of molecular sciences

|August 14, 2025

概括

这项研究提出了一种新的方法,用于使用单个DNA磁带共同表达指核酶 (ZFN) 单体. 这种方法减少了DNA大小,增强了ZFN在基因组编辑的病毒载体中的应用.

科学领域:

分子生物学分子生物学
基因编辑技术的技术
生物技术是生物技术.

背景情况:

基因组编辑是生物医学研究中的关键工具.
指核酶 (ZFNs) 比其他基因组编辑工具 (如TALENs和CRISPR-Cas9) 更小,因此它们适合病毒载体.
一个ZFN的局限性是需要分别表达左和右单体,需要两个DNA磁带,这增加了矢量大小.

研究的目的:

开发一种从单个表达盒中共同表达ZFN单体的方法.
为了克服ZFN传递病毒载体的尺寸限制.
为了保持或提高基因组编辑效率,使用新的共同表达系统.

主要方法:

从单个DNA磁带中进行联合表达的工程T2A合的ZF-ND1单体.
单个表达卡塞特转化为细胞.
评估了DNA裂变效率,并将基因组编辑结果与两卡塞特系统进行了比较.

主要成果:

通过使用单个T2A合表达盒成功证明了ZF-ND1单体的联合表达.
同表达的ZF-ND1s有效地切割了目标DNA序列.
与两个单独的单体相比,减少了总被转染的等离子体DNA量的一半,同时实现了与两个单独的单体相比相当的基因组编辑效率.

结论:

通过单个T2A合磁带共同表达ZFN单体是可行的和高效的.
这一策略显著减少了DNA有效载荷大小,有利于病毒载体应用.
为推进基于ZFN的基因组编辑技术提供了一个有前途的框架.

关键词:

这是一种2A.ZFNN 在线播放基因组编辑基因组编辑

更多相关视频

Mouse Genome Engineering Using Designer Nucleases

Mouse Genome Engineering Using Designer Nucleases

Published on: April 2, 2014

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

Published on: December 11, 2020

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...

Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

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...