Analyte-induced hindrance in the RCA-assisted CRISPR/Cas12a system for homogeneous protein assays

Mao，Guobin1; Li，Qiaoyu1; Zhang，Ziying1; Huang，Wei5; Luo，Qian3; Dai，Junbiao1,2; Huang，Weiren1,4; Ma，Yingxin1

2024-11-22

10.1016/j.aca.2024.343294

Analytica Chimica Acta   影响因子和分区

0003-2670

1873-4324

Heterogeneous assays, such as enzyme-linked immunosorbent assays, have become indispensable for in vitro diagnostics. However, the simple, sensitive, and accurate detection is limited by their multiple washing and incubation steps, and limited amplification methods. In this study, we design a novel approach utilizing analyte-induced hindrance within the rolling circle amplification (RCA)-assisted CRISPR/Cas12a system for simple and highly sensitive homogenous protein detection. Streptavidin (SA) and digoxin antibody (anti-Dig) are employed as representative detection models. The specific recognition of target proteins using primers modified with small molecules hinders the RCA process, preventing the activation of Cas12a′s trans-cleavage activity, thereby leading to a reduction in fluorescence intensity. Our developed platform exhibites exceptional detection performance characterized by high sensitivity, robust specificity, and significant potential for application in complex samples. By expanding the recognition elements, this platform can evolve into a versatile clinical diagnostic tool with universal applicability. In addition, this platform provides a novel direction for quantifying ultralow-concentration disease biomarkers in clinical practice.

Biosensing CRISPR/Cas12a Homogeneous assay Protein detection RCA

英语

其他

2-s2.0-85205305899

Scopus

1.CAS Key Laboratory of Quantitative Engineering Biology,Guangdong Provincial Key Laboratory of Synthetic Genomics and Shenzhen Key Laboratory of Synthetic Genomics,Shenzhen Institute of Synthetic Biology,Shenzhen Institutes of Advanced Technology,Chinese Academy of Sciences,Shenzhen,518055,China
2.Shenzhen Branch,Guangdong Laboratory of Lingnan Modern Agriculture,Key Laboratory of Synthetic Biology,Ministry of Agriculture and Rural Affairs,Agricultural Genomics Institute at Shenzhen,Chinese Academy of Agricultural Sciences,Shenzhen,518000,China
3.Shenzhen Institutes of Advanced Technology,Chinese Academy of Sciences,Shenzhen,518055,China
4.Department of Urology,Shenzhen Institute of Translational Medicine,Shenzhen Second People's Hospital,The First Affiliated Hospital of Shenzhen University,International Cancer Center,Shenzhen University School of Medicine,Shenzhen,518039,China
5.Department of Biology,Southern University of Science and Technology,Shenzhen,518055,China

Mao，Guobin,Li，Qiaoyu,Zhang，Ziying,et al. Analyte-induced hindrance in the RCA-assisted CRISPR/Cas12a system for homogeneous protein assays[J]. Analytica Chimica Acta,2024,1330.

Mao，Guobin.,Li，Qiaoyu.,Zhang，Ziying.,Huang，Wei.,Luo，Qian.,...&Ma，Yingxin.(2024).Analyte-induced hindrance in the RCA-assisted CRISPR/Cas12a system for homogeneous protein assays.Analytica Chimica Acta,1330.

Mao，Guobin,et al."Analyte-induced hindrance in the RCA-assisted CRISPR/Cas12a system for homogeneous protein assays".Analytica Chimica Acta 1330(2024).