Y-shaped DNA nanostructures assembled-spherical nucleic acids as target converters to activate CRISPR-Cas12a enabling sensitive ECL biosensing

作者全名："-Ling, Liu Mei; Yi, Li; -Ling, Zhao Mei; Ying, Zhuo; Xiao-Jing, He"

作者地址："[-Ling, Liu Mei; Yi, Li; Xiao-Jing, He] Chongqing Med Univ, Affiliated Hosp 2, Dept Radiol, Chongqing, Peoples R China; [-Ling, Zhao Mei; Ying, Zhuo] Southwest Univ, Coll Chem & Chem Engn, Key Lab Luminescence Anal & Mol Sensing, Minist Educ, Chongqing, Peoples R China"

通信作者："Xiao-Jing, H (通讯作者)，Chongqing Med Univ, Affiliated Hosp 2, Dept Radiol, Chongqing, Peoples R China.; Ying, Z (通讯作者)，Southwest Univ, Coll Chem & Chem Engn, Key Lab Luminescence Anal & Mol Sensing, Minist Educ, Chongqing, Peoples R China."

来源：BIOSENSORS & BIOELECTRONICS

ESI学科分类：CHEMISTRY

WOS号：WOS:000826361500001

JCR分区：Q1

影响因子：12.6

年份：2022

卷号：214

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：　

摘要："Considering the trans-cleavage capabilities, high-specificity and programmability, the CRISPR-Cas system has been recognized as a valuable platform to develop the next-generation diagnostic biosensors. However, due to the natural interaction with nucleic acids, current CRISPR-Cas-based detection mostly applies in nucleic acid analysis rather than non-nucleic acid analysis. By virtue of spherical nucleic acids (SNAs) with programmability and specificity, the Y-shaped DNA nanostructures assembled-SNAs (Y-SNAs) were rationally designed as target converters to achieve the quantitative activation of CRISPR-Cas12a, enabling a highly specific and sensitive electrochemiluminescence (ECL) determination of alpha-methylacyl-CoA racemase (AMACR), a high specific protein biomarker of prostate cancer. Significantly, the Y-shaped DNA nanostructures comprised of assisted DNA (A1), AMACR aptamer and DNA activator of CRISPR-Cas12a were loaded on Au nanoparticles modified Fe3O4 magnetic beads (Au@Fe3O4 MBs) to construct the robust Y-SNAs. In the presence of the target AMACR, the Y-SNAs as target converters could achieve quantitative activation of CRISPR-Cas12a by outputting the DNA activators with a linear relationship to the target. The amplified ECL signals were triggered by the release of the ferrocene-labeled quenching probes (QPs) on the electrode surface due to the trans-cleavage activity of CRISPR-Cas12a, thereby realizing the sensitive ECL determination of AMACR from 10 ng/mL to 100 mu g/mL with the detection limit of 1.25 ng/mL. In general, this approach provides novel perspectives on how to design a universal ECL platform of the CRISPR-Cas system to detect the non-nucleic acid targets beyond the traditional methods."

基金机构："General program of Chongqing Natural Science Foundation [cstc2019jcyj-msxmX0073]; Science and Health Joint Medical Research Project of Chongqing [2019GDRC011]; Senior Medical Talents program of Chongqing for Young and Middle- aged, and Kuanren Talents Program of the second affiliated hospital of Chongqing Medical University"

基金资助正文："Data availability I have shared the link to my date at the Attach File step. Acknowledgments This work was supported by the General program of Chongqing Natural Science Foundation (cstc2019jcyj-msxmX0073) , Science and Health Joint Medical Research Project of Chongqing (2019GDRC011) , Senior Medical Talents program of Chongqing for Young and Middle- aged, and Kuanren Talents Program of the second affiliated hospital of Chongqing Medical University."