相关实验视频

Updated: Jul 7, 2025

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

Published on: August 25, 2017

基因点突变信息的翻译和检测:利用单基扩展和CRISPR/Cas12a

Zhujun Liu¹, Jie Xu¹, Shan Huang¹

¹School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.

Biosensors & bioelectronics

|December 24, 2023

概括

这项研究提出了一种新的CRISPR/Cas12a方法来检测基因点突变,这对于癌症诊断和治疗至关重要. 这种增强技术为改善临床癌症查提供了高灵敏度,特异性和通用性.

科学领域:

分子生物学分子生物学
遗传学是一个遗传学.
生物技术是生物技术.

背景情况:

基因点突变是癌症发展的关键驱动因素.
准确检测这些突变对于临床应用至关重要.
目前用于突变检测的CRISPR/Cas12a方法在普遍性和特异性方面存在局限性.

研究的目的:

开发一种基于CRISPR/Cas12a的普遍适用,敏感和特定的基因点突变检测方法.
克服现有的CRISPR/Cas12a系统的局限性,特别是PAM序列依赖性.
为了将DNA点突变信息转化为可检测的长度变化.

主要方法:

开发了一个CRISPR/Cas12a检测系统.
综合改进的基因特异性聚合酶链反应 (PCR) 和单基扩展.
将目标双链DNA (dsDNA) 点突变信息翻译成单链DNA (ssDNA) 激活器长度变化.
克服了原始空间体相邻动机 (PAM) 序列限制.

主要成果:

对于EGFR T790M突变,已达到0.002%的检测极限.
在检测基因点突变方面表现出高灵敏度和特异性.
构建的CRISPR/Cas12a系统显示了广泛的适用性 (普遍性).

结论:

新的CRISPR/Cas12a方法有效地检测出具有高灵敏度和特异性的基因点突变.
这种方法克服了以前基于CRISPR的检测系统的局限性.
开发的方法是临床癌症查和个性化医学的一个有前途的工具.

关键词:

基特异性聚合酶连锁反应这就是CRISPR/Cas12a.基因点突变基因点突变信息翻译信息翻译单个基础扩展单个基础扩展

更多相关视频

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

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

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...

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...