相关实验视频

Updated: Mar 22, 2026

Generation of Defined Genomic Modifications Using CRISPR-CAS9 in Human Pluripotent Stem Cells

Generation of Defined Genomic Modifications Using CRISPR-CAS9 in Human Pluripotent Stem Cells

Published on: September 25, 2019

使用CRISPR/Cas9有效地引入特定的同位素和异位素突变

Dominik Paquet¹, Dylan Kwart¹, Antonia Chen¹

¹Laboratory of Brain Development and Repair, The Rockefeller University, 1230 York Avenue, New York, New York 10065, USA.

|April 28, 2016

概括

这项研究引入了用于精确基因修改的CRISPR/Cas9基因组编辑框架. CORRECT方法提高了同质导向修复 (HDR) 的效率,使得阿尔茨海默氏症等遗传疾病的准确建模成为可能.

科学领域:

分子生物学
遗传学
生物技术

背景情况:

CRISPR/Cas9能够进行基因编辑,但对精确突变的同源导向修复 (HDR) 是无效的.
目前的方法在疾病建模中难以针对异构基因突变.
不同类末端连接 (NHEJ) 往往会引入不需要的插入/删除.

研究的目的:

开发一种高效准确的CRISPR/Cas9基因组编辑框架,用于选择性单基因和双基因序列改变.
提高基因疾病建模的同质导向修复 (HDR) 的效率和准确性.
为了使单个等位基因的向突变敲击能够模拟异构基因突变.

主要方法:

开发了一个包含静音CRISPR/Cas9阻断突变的CRISPR/Cas9基因组编辑框架.
建立了一个名为"CORRECT"的无痕基因组编辑方法.
利用突变结合率与双链断裂 (DSB) 之间的关系来控制结合性.

主要成果:

实现了单基和双基序列变化的高效和精确引入.
通过结合静音阻断突变, 显著提高了HDR准确性.
成功产生具有异构和同构阿尔茨海默病突变的人类诱导多能干细胞 (APP,PSEN1和M146V).

结论:

开发的框架和CORRECT方法可使用CRISPR/Cas9进行高效和精确的基因编辑.
这种方法可用于精确的疾病建模.
通过有效引入特定序列变化,促进人类疾病的研究.

更多相关视频

Construction of Homozygous Mutants of Migratory Locust Using CRISPR/Cas9 Technology

Construction of Homozygous Mutants of Migratory Locust Using CRISPR/Cas9 Technology

Published on: March 16, 2022

Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format

Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format

Published on: April 8, 2017

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...

CRISPR

CRISPR

Homologous Recombination

Homologous Recombination

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...

CRISPR and crRNAs

CRISPR and crRNAs

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...