清脆的
生物传感器
DNA
化学
滚动圆复制
核酸
互补DNA
连接器
组合化学
生物物理学
分子生物学
生物化学
聚合酶
生物
基因
计算机科学
操作系统
作者
Mengmeng Chen,Jingyang Zhang,Yuan Peng,Jialei Bai,Shuang Li,Dianpeng Han,Shuyue Ren,Kang Qin,Huanying Zhou,Tie Han,Yu Wang,Zhixian Gao
标识
DOI:10.1016/j.bios.2022.114792
摘要
Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas systems exhibit significant potential in developing biosensing technology due to their collateral cleavage capabilities. Herein, we introduced the collateral cleavage activity of CRISPR/Cas14a to activate DNA hydrogel for ultrasensitive detection of the myocardial infarction biomarker creatine kinase MB (CK-MB). In this strategy, the designed CRISPR/Cas14a system can be activated by introducing complementary DNA (cDNA) derived from competitive dissociation and exponential amplification (EXPAR), which is positively correlated with creatine kinase isoenzyme (CK-MB) concentration. Then the activated Cas14a protein can be utilized to indiscriminately cleave the DNA hydrogel cross-linker strand, leading to the degradation of the gel matrix and thus releasing the pre-encapsulated PtNPs/Cu-TCPP(Fe). PtNPs/Cu-TCPP(Fe) can trigger the TMB reaction, leading to an increase in absorbance value at 450 nm, thus enabling the quantitative detection of CK-MB. The proposed strategy combines CRISPR/Cas14a with DNA hydrogel for the first time, improving the programmability of DNA hydrogel and providing a reliable, sensitive, and versatile detection platform for trace non-nucleic acid targets.
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