Liposome fusogenic enzyme-free circuit enables high-fidelity determination of single exosomal RNA

Zixin Sun; Xiaohui Chen; Ruyan Niu; Hengyi Chen; Ying Zhu; Chong Zhang; Liu Wang; Huaming Mou; Hong Zhang; Yang Luo

doi:10.1016/j.mtbio.2023.100613

Liposome fusogenic enzyme-free circuit enables high-fidelity determination of single exosomal RNA

Mater Today Bio. 2023 Mar 20:19:100613. doi: 10.1016/j.mtbio.2023.100613. eCollection 2023 Apr.

Authors

Zixin Sun^{1

2}, Xiaohui Chen^{1

3}, Ruyan Niu^{1

3}, Hengyi Chen¹, Ying Zhu¹, Chong Zhang^{1

3}, Liu Wang¹, Huaming Mou², Hong Zhang⁴, Yang Luo¹

Affiliations

¹ Department of Center of Smart Laboratory and Molecular Medicine, School of Medicine, Chongqing University, Chongqing, 400044, PR China.
² Sanxia Hospital, Chongqing University, Chongqing, 404100, PR China.
³ Key Laboratory for Biotechnological Science and Technology of Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, 400044, PR China.
⁴ Department of Clinical Laboratory, The Second Hospital of Shandong University, Jinan, Shandong, 250033, PR China.

Abstract

Accurate determination of single exosomal inclusions in situ presents a significant challenge due to their extremely low abundance as well sub-100 nm vesicle dimensions. Here, we created a Liposome Fusogenic Enzyme-free circuit (LIFE) approach for the high-fidelity identification of exosome-encapsulated cargoes without destroying the vesicle integrity. The probe-loaded cationic fusogenic liposome could capture and fuse with a single target exosome, enabling probes delivery and target biomolecule-initiated cascaded signal amplification in situ. Then the DNAzyme probe encountered conformal change upon exosomal microRNA activation, and generated a convex DNAzyme structure to cleave the RNA site of substrate probe. After that, the target microRNA could be released to introduce a cleavage cycle to yield amplified fluorescence readout. Therefore, trace cargoes in a single exosome could be accurately determined by elaborately controlling the ratio of introduced LIFE probe, paving the way toward the exploration of a universal sensing platform for the assessment of exosomal cargoes to facilitate early disease diagnosis and personalized treatment.

Keywords: Exosome; Fusion; Liposome; Single-molecule detection; Tumor.