相关实验视频
Updated: May 22, 2025
08:50
A Nonsequencing Approach for the Rapid Detection of RNA Editing
Published on: April 21, 2022
2.5K
基于TALE的C-to-T基基编辑器用于多个同类基因,具有灵活的精度
Ayako Hosoda1, Issei Nakazato1,2, Miki Okuno3
1Laboratory of Plant Molecular Genetics, Graduate School of Agricultural and Life Science, The University of Tokyo, Tokyo 113-8657, Japan.
Plant biotechnology (Tokyo, Japan)
|March 14, 2025
概括
一种新的基于TALE的基准编辑方法可以在Arabidopsis中进行精确的C-to-T替代. 这种方法扩大了向范围,并减少了基因组工程的非向突变.
科学领域:
- 分子生物学分子生物学
- 遗传学 遗传学 是一个
- 植物科学 植物科学
背景情况:
- 克里斯普尔-Cas9系统提供高效的C-to-T基编辑,但在准范围和非目标效应方面存在局限性.
- 基于转录激活器样效应器 (TALE) 的工具提供了更长的DNA识别序列,为增强的基编辑特异性提供了潜在的潜力.
研究的目的:
- 在Arabidopsis中开发一种新的基编辑系统,使用与TALE DNA结合域合的cytidine deaminase.
- 通过扩大准范围和最大限度地减少非准突变来克服CRISPR介导的基准编辑的局限性.
主要方法:
- 在Arabidopsis核基因中,将细胞丁除氨酶融合到TALE DNA结合域,用于针对性的C-to-T基基编辑.
- 利用一种能够识别所有四个DNA基的新型TALE重复单元,使得可变的编码子位置的向成为可能.
- 采用单一的TALE对用于同时编辑多个基因异型.
主要成果:
- 在Arabidopsis核基因中实现了高效和有针对性的C-to-T替代.
- 与以前的方法相比,基于TALE的系统显示了扩大目标范围.
- 观察到抑制了目标之外的替换,提高了编辑精度.
- 成功地证明了对多个非常相似的基因异型的同时基因编辑.
结论:
- 开发的TALE-cytidine deaminase融合系统提供了一种高效和精确的方法,用于Arabidopsis中C-to-T基编辑.
- 这一策略通过提供更广泛的准和更好的特异性来增强基因组工程能力,特别是在复杂的基因家族中.
相关概念视频
02:33Combinatorial Gene Control
8.3K
Combinatorial gene control is the synergistic action of several transcriptional factors to regulate the expression of a single gene. The absence of one or more of these factors may lead to a significant difference in the level of gene expression or repression.
The expression of more than 30,000 genes is controlled by approximately 2000-3000 transcription factors. This is possible because a single transcription factor can recognize more than one regulatory sequence. The specificity in gene...
The expression of more than 30,000 genes is controlled by approximately 2000-3000 transcription factors. This is possible because a single transcription factor can recognize more than one regulatory sequence. The specificity in gene...
8.3K
02:02Cis-regulatory Sequences
2.9K
2.9K
02:53Conservative Site-specific Recombination and Phase Variation
5.9K
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...
5.9K
02:27Base-pairing and DNA Repair
64.5K
64.5K
02:31Homologous Recombination
50.0K
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...
50.0K
03:36Genome Annotation and Assembly
18.8K
The genome refers to all of the genetic material in an organism. It can range from a few million base pairs in microbial cells to several billion base pairs in many eukaryotic organisms. Genome assembly refers to the process of taking the DNA sequencing data and putting it all back together in a correct order to create a close representation of the original genome. This is followed by the identification of functional elements on the newly assembled genome, a process called genome annotation.
18.8K