Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Dual-mode colorimetric and fluorescence detection of BRCA1 based on a CRISPR-Cas12a system
Analyst ( IF 3.6 ) Pub Date : 2024-08-08 , DOI: 10.1039/d4an01035c Chengchen Tan 1 , Xiaolong Yan 1 , Xingchang Lu 1 , Jianxiu Wang 1 , Xinyao Yi 1
Analyst ( IF 3.6 ) Pub Date : 2024-08-08 , DOI: 10.1039/d4an01035c Chengchen Tan 1 , Xiaolong Yan 1 , Xingchang Lu 1 , Jianxiu Wang 1 , Xinyao Yi 1
Affiliation
|
Breast cancer, the most common malignant tumor in the world, seriously threatens human life and health. Early diagnosis of breast cancer may help enhance the survival rate. In this work, a colorimetric and fluorescent dual-mode biosensor based on the CRISPR-Cas12a system was constructed to detect the breast cancer biomarker BRCA1. The intact G4 DNA, with the assistance of K+ and hemin, catalyses the oxidation of o-phenylenediamine (OPD) with the assistance of hydrogen peroxide (H2O2), generating the oxidation product 2,3-diaminophenazine (DAP), which has distinct absorption and fluorescence peaks. The presence of the target BRCA1 activates the trans-cleavage activity of CRISPR-Cas12a, leading to the cleavage of G4 DNA and inhibiting the catalytic oxidation of OPD. Target BRCA1 was quantitatively determined by measuring both the absorbance and fluorescence intensity of DAP. The detection limits were calculated to be 0.615 nM for the colorimetric method and 0.289 nM for the fluorescence method. The dual-mode biosensor showed good selectivity and reliability for BRCA1 and can resist interference from complex substrates, and it has great potential in biomedical detection.
中文翻译：
基于CRISPR-Cas12a系统的BRCA1双模式比色和荧光检测
乳腺癌是世界上最常见的恶性肿瘤，严重威胁人类的生命和健康。乳腺癌的早期诊断可能有助于提高生存率。本工作构建了基于CRISPR-Cas12a系统的比色荧光双模式生物传感器来检测乳腺癌生物标志物BRCA1。完整的G4 DNA在K +和氯高铁血红素的帮助下，在过氧化氢（H 2 O 2 ）的帮助下催化邻苯二胺（OPD）的氧化，生成氧化产物2,3-二氨基吩嗪（DAP），具有明显的吸收峰和荧光峰。靶标BRCA1的存在会激活CRISPR-Cas12a的反式切割活性，导致G4 DNA的切割并抑制OPD的催化氧化。通过测量 DAP 的吸光度和荧光强度来定量确定目标 BRCA1。计算出比色法的检测限为 0.615 nM，荧光法的检测限为 0.289 nM。该双模式生物传感器对BRCA1表现出良好的选择性和可靠性，并且能够抵抗复杂底物的干扰，在生物医学检测方面具有巨大的潜力。
更新日期：2024-08-08
中文翻译：
基于CRISPR-Cas12a系统的BRCA1双模式比色和荧光检测
乳腺癌是世界上最常见的恶性肿瘤，严重威胁人类的生命和健康。乳腺癌的早期诊断可能有助于提高生存率。本工作构建了基于CRISPR-Cas12a系统的比色荧光双模式生物传感器来检测乳腺癌生物标志物BRCA1。完整的G4 DNA在K +和氯高铁血红素的帮助下，在过氧化氢（H 2 O 2 ）的帮助下催化邻苯二胺（OPD）的氧化，生成氧化产物2,3-二氨基吩嗪（DAP），具有明显的吸收峰和荧光峰。靶标BRCA1的存在会激活CRISPR-Cas12a的反式切割活性，导致G4 DNA的切割并抑制OPD的催化氧化。通过测量 DAP 的吸光度和荧光强度来定量确定目标 BRCA1。计算出比色法的检测限为 0.615 nM，荧光法的检测限为 0.289 nM。该双模式生物传感器对BRCA1表现出良好的选择性和可靠性，并且能够抵抗复杂底物的干扰，在生物医学检测方面具有巨大的潜力。
京公网安备 11010802027423号