Rapid and accurate genotyping of human SNP rs671 in aldehyde dehydrogenase 2 gene using one-step CRISPR/Cas12b assay without DNA amplification.

IF 2.8 4区 生物学 Q3 CELL BIOLOGY Cell Division Pub Date : 2023-08-28 DOI:10.1186/s13008-023-00095-6
Fang Wu, Yong Xue, Yan Wang, Xinxin Si, Xinyue Zhang, Yuyang Xu, Zhidan Luo
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Abstract

Background: The SNP rs671 of Human aldehyde dehydrogenase (ALDH) is G-A transition at 1510th nucleotides, which is an important clinical indicator of alcoholic liver disease, digestive tract cancer and some drug efficiency. The commonly used genotyping assay of this polymorphism is relatively time-consuming and costly.

Finding: This study develops a rapid and accurate one-step CRISPR/Cas12b assay to distinguish the G1510A polymorphism of human ALDH2 free of DNA amplification. The method we established requires only one step of adding 1 μl genomic DNA sample to premixed system, and waiting for the acquisition of fluorescent signal, taking approximate 30 min.

Conclusions: This method provides a potential tool for more accurate and reliable nucleic acid detection with a single base difference and supports the relevant disease diagnosis and personalized medicine.

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无需DNA扩增,利用CRISPR/Cas12b一步法快速准确分型人醛脱氢酶2基因SNP rs671
背景:人醛脱氢酶(Human aldehyde dehydrogenase, ALDH) SNP rs671在1510个核苷酸处发生G-A转位,是酒精性肝病、消化道肿瘤及部分药物疗效的重要临床指标。这种多态性的常用基因分型分析相对耗时和昂贵。发现:本研究建立了一种快速准确的一步CRISPR/Cas12b检测方法,用于区分无DNA扩增的人ALDH2基因G1510A多态性。所建立的方法只需要将1 μl基因组DNA样品加入到预混体系中,等待荧光信号的获取,耗时约30 min。结论:该方法为更准确可靠的单碱基差核酸检测提供了潜在的工具,为相关疾病诊断和个性化医疗提供了支持。
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来源期刊
Cell Division
Cell Division CELL BIOLOGY-
CiteScore
3.70
自引率
0.00%
发文量
5
审稿时长
>12 weeks
期刊介绍: Cell Division is an open access, peer-reviewed journal that encompasses all the molecular aspects of cell cycle control and cancer, cell growth, proliferation, survival, differentiation, signalling, gene transcription, protein synthesis, genome integrity, chromosome stability, centrosome duplication, DNA damage and DNA repair. Cell Division provides an online forum for the cell-cycle community that aims to publish articles on all exciting aspects of cell-cycle research and to bridge the gap between models of cell cycle regulation, development, and cancer biology. This forum is driven by specialized and timely research articles, reviews and commentaries focused on this fast moving field, providing an invaluable tool for cell-cycle biologists. Cell Division publishes articles in areas which includes, but not limited to: DNA replication, cell fate decisions, cell cycle & development Cell proliferation, mitosis, spindle assembly checkpoint, ubiquitin mediated degradation DNA damage & repair Apoptosis & cell death
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