相关实验视频

Updated: May 1, 2026

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

在合成DNA,细胞系cfDNA模型和液体活检上基于CRISPR-Cas12a的高精度SNV检测协议

Kavish A V Kohabir¹, Jasper Linthorst², Rob M F Wolthuis³

¹Department of Human Genetics, Amsterdam UMC Location Vrije Universiteit Amsterdam, Amsterdam, the Netherlands; Amsterdam Reproduction & Development, Amsterdam, the Netherlands; Imaging and Biomarkers, Cancer Center Amsterdam, Amsterdam, the Netherlands.

|March 15, 2025

概括

这项研究引入了基于Cas12a的CRISPR诊断 (CRISPRdx) 方法,用于高精度的单核酸变体检测. 该协议提高了使用无细胞DNA和液体活检的基因测试的精度.

科学领域:

分子生物学分子生物学
遗传学遗传学是一个
生物技术是生物技术.

背景情况:

基于CRISPR的诊断 (CRISPRdx) 显示出快速,负担得起的基因测试的潜力.
在CRISPRdx分析中实现单核酸分辨率仍然是一个重要的技术障碍.

研究的目的:

提出使用基于Cas12a的系统高精度检测单核酸变异 (SNV) 的协议.
展示ARTEMIS算法用于SNV识别和CRISPRRNA (crRNA) 设计的应用.

主要方法:

使用基于Cas12a的CRISPRdx方法进行SNV检测.
采用ARTEMIS算法用于目标SNV识别和crRNA设计.
在合成DNA,细胞系衍生的无细胞DNA (cfDNA) 和液体活检样本上进行了基于光的CRISPRdx测定.

主要成果:

开发了一种允许高精度检测SNVs的协议.
成功地将该方法应用于各种样本类型,包括cfDNA和液体活检.
证明了ARTEMIS算法在优化CRISPRdx试验中的实用性.

结论:

基于Cas12a的协议提供了一个精确的SNV检测方法,解决了CRISPRdx.的关键局限性.
这种方法可以使用随时可用的生物样本进行准确的基因测试.
描述的协议和算法为推进遗传诊断提供了有价值的工具.

关键词:

克里斯普尔是什么意思?克里斯普尔是什么意思?生物技术和生物工程生物技术和生物工程癌症癌症癌症癌症癌症细胞生物学细胞生物学细胞培养培养的细胞培养.遗传学遗传学遗传学是一个

更多相关视频

Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens

Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens

Published on: June 6, 2020

DNA Virus Detection System Based on RPA-CRISPR/Cas12a-SPM and Deep Learning

DNA Virus Detection System Based on RPA-CRISPR/Cas12a-SPM and Deep Learning

Published on: May 10, 2024

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...

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...

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...