相关实验视频
Updated: May 23, 2025
07:56
Genome Editing in Mammalian Cell Lines using CRISPR-Cas
Published on: April 11, 2019
21.5K
CrisprBuildr是一个开源的应用程序,用于在Drosophila melanogaster中使用CRISPR介导的基因组工程
Nicole Horsley1,2, Adam von Barnau Sythoff1,3, Mark Delgado4
1Department of Biology, University of Washington, Life Science Building, Seattle WA 98195, USA.
bioRxiv : the preprint server for biology
|March 10, 2025
概括
CrisprBuildr通过启用基因删除或光蛋白标记来简化CRISPR/Cas9基因组工程. 这种开源工具有助于研究人员设计指导RNA和修复模板以进行体内蛋白质表征.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 生物技术是生物技术.
背景情况:
- 克里斯普尔/卡斯9技术使得基因改造和蛋白质研究的向基因工程成为可能.
- 为CRISPR/Cas9实验设计和克隆构造通常是耗时和劳动密集的.
- 需要有效的工具来简化基因删除和蛋白质标记的过程.
研究的目的:
- 开发一个基于Web的应用程序CrisprBuildr,以简化基于CRISPR/Cas9的基因组工程.
- 为了促进基因的删除或插入光蛋白 (例如,EGFP,mCherry) 在特定的蛋白质末端.
- 为设计指导RNA和修复模板向量提供一个用户友好的平台.
主要方法:
- 开发CrisprBuildr,这是一个开源的,基于Web的应用程序.
- 整合Drosophila melanogaster基因组作为主要模板,可适应其他基因组.
- 为基于iLID的光遗传学,生成包含光蛋白和SspB-Q73R的新型标记载体.
主要成果:
- CrisprBuildr使用户能够设计基因删除或N-/C-终端光蛋白插入的结构.
- 该应用程序通过指导RNA和修复模板向量的设计指导用户.
- 为EGFP,mCherry和光遗传学应用生成了新的标记载体.
结论:
- CrisprBuildr显著减少了CRISPR/Cas9构造设计所需的时间和精力.
- 该工具支持有效的蛋白质标记和基因操纵,用于体内研究.
- 未来的开发将扩大CrisprBuildr的功能,提供更多的矢量,基因组和CRISPR应用程序.
相关概念视频
02:53CRISPR and crRNAs
16.5K
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...
16.5K
01:59CRISPR
48.8K
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...
48.8K
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
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