Multiplex SNP Genotyping Using SWITCH: Sequence‐Specific Nanoparticle with Interpretative Toehold‐Mediated Sequence Decoding in Hydrogel

• Published in: Small (Weinheim an der Bergstrasse, Germany) Vol. 18; no. 8; pp. e2105538 - n/a
• Main Authors: Choi, Woongsun, Park, Eunhye, Bae, Seojin, Choi, Kyung‐Hak, Han, Sangeun, Son, Kuk‐Hui, Lee, Do Young, Cho, Il‐Joo, Seong, Hyejeong, Hwang, Kyo Seon, Nam, Jwa‐Min, Choi, Jungkyu, Lee, Hyojin, Choi, Nakwon
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 01.02.2022
• Subjects: Alleles; Biological properties; Fluorescence; Genotype; Gold; gold nanoparticles; hydrogel microparticles; Hydrogels; magnetic microparticles; Metal Nanoparticles; Microparticles; multiplex single nucleotide polymorphism genotyping; Multiplexing; Nanoparticles; Nanotechnology; Nucleotides; Polymerase chain reaction; Polymorphism, Single Nucleotide; sandwich assays; toehold‐mediated DNA displacement; warfarin; hydrogel microparticles; magnetic microparticles; gold nanoparticles; toehold-mediated DNA displacement; multiplex single nucleotide polymorphism genotyping; sandwich assays; warfarin
• ISSN: 1613-6810; 1613-6829
• DOI: 10.1002/smll.202105538

Single nucleotide polymorphisms (SNPs) that can alter phenotypes of individuals play a pivotal role in disease development and, more importantly, responses to therapy. However, SNP genotyping has been challenging due to the similarity of SNP alleles and their low concentration in biological samples. Sequence‐specific nanoparticle with interpretative toehold‐mediated sequence decoding in hydrogel (SWITCH) for multiplex SNP genotyping is presented. The encoding with gold nanoparticle probes transduces each SNP target to ≈1000 invaders with prominently different sequences between wild and mutant types, featuring polymerase chain reaction (PCR)‐free amplification. Subsequently, the toehold‐mediated DNA replacement in hydrogel microparticles decodes the invaders via SNP‐specific fluorescence signals. The 4‐plex detection of the warfarin‐associated SNP targets spiked in commercially validated human serum (S1‐100ML, Merck) is successfully demonstrated with excellent specificity. This work is the first technology development presenting PCR‐free, multiplex SNP genotyping with a single reporting fluorophore, to the best of knowledge. The SWITCH is the first development presenting polymerase chain reaction‐free, multiplex single nucleotide polymorphism genotyping with a single reporting fluorophore by a hybrid assay using inorganic and organic materials.