DataSheet2_CRISPR/Cas12a-Assisted Visual Logic-Gate Detection of Pathogenic Microorganisms Based on Water-Soluble DNA-Binding AIEgens.docx

Here, we developed a rapid, visual and double-checked Logic Gate detection platform for detection of pathogenic microorganisms by aggregation-induced emission luminogens (AIEgens) in combination with Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR associated (Cas). DNA light-up AIEgens (1,1,2,2-tetrakis[4-(2-bromo-ethoxy) phenyl]ethene, TTAPE) was non-emissive but the emission was turned on in the presence of large amount of DNA produced by recombinase polymerase amplification (RPA). When CRISPR/Cas12a was added, all long-stranded DNA were cut leading to the emission quenched. Thus, a method that can directly observe the emission changes with the naked eye has been successfully constructed. The detection is speedy within only 20 min, and has strong specificity to the target. The result can be judged by Logic Gate. Only when the output signal is (1,0), does it represent the presence of pathogenic microorganisms in the test object. Finally, the method was applied to the detect pathogenic microorganisms in environmental water samples, which proved that this method has high selectivity, specificity and applicability for the detection of pathogenic microorganisms in environmental water samples.

本研究团队构建了一项快速、直观且经过双重验证的逻辑门检测平台，用于通过聚诱导发光发光原体（AIEgens）检测致病微生物，该平台结合了成簇规律间隔短回文重复序列（CRISPR）/CRISPR相关（Cas）技术。DNA发光AIEgens（1,1,2,2-四[4-(2-溴乙氧基)苯基]乙炔，TTAPE）在无DNA存在时无发光现象，而当重组酶聚合酶扩增（RPA）产生大量DNA时，发光得以激活。加入CRISPR/Cas12a后，所有长链DNA被切割，导致发光淬灭。因此，一种能够通过肉眼直接观察发光变化的方法得以成功构建。检测过程仅需20分钟，对目标具有高度特异性。结果可通过逻辑门进行判断，只有当输出信号为（1，0）时，才表明测试样本中存在致病微生物。最终，该方法被应用于检测环境水样中的致病微生物，证明该方法在环境水样中致病微生物的检测方面具有较高的选择性、特异性和适用性。