Characterization and application of fluidic properties of trinucleotide repeat sequences by wax-on-plastic microfluidics. Issue 4 (2nd January 2020)

• Record Type: Journal Article
• Authors: Qamar, Ahmad Zaman; Asefifeyzabadi, Narges; Taki, Motahareh; Naphade, Swati; Ellerby, Lisa M.; Shamsi, Mohtashim Hassan
• Is Part Of: Journal of materials chemistry. Volume 8:Issue 4(2019)
• Page Start: 743
• Page End: 751
• Publication Date: 2020-01-02
• Subjects: Materials -- Periodicals; Chemistry, Analytic -- Periodicals; Biomedical materials -- Research -- Periodicals; 543.0284
• DOI: 10.1039/c9tb02208b
• Languages: English
• ISSNs: 2050-750X

Abstract

Abstract : This work demonstrates sequence directed flexibility of neurodegenerative trinucleotide repeat sequences can be translated into distinct capillary flow using wax-on-plastic microfluidics and used to detect microsatellite repeat expansions diseases. Abstract : Trinucleotide repeat (TNR) sequences introduce sequence-directed flexibility in the genomic makeup of all living species leading to unique non-canonical structure formation. In humans, the expansions of TNR sequences are responsible for almost 24 neurodegenerative and neuromuscular diseases because their unique structures disrupt cell functions. The biophysical studies of these sequences affect their electrophoretic mobility and spectroscopic signatures. Here, we demonstrate a novel strategy to characterize and discriminate the TNR sequences by monitoring their capillary flow in the absence of an external driving force using wax-on-plastic microchannels. The wax-on-plastic microfluidic system translates the sequence-directed flexibility of TNR into differential flow dynamics. Several variables were used to characterize sequences including concentration, single- vs. double-stranded samples, type of repeat sequence, length of the repeat sequence, presence of mismatches in duplex, and presence of metal ion. All these variables were found to influence the flow velocities of TNR sequences as these factors directly affect the structural flexibility of TNR at the molecular level. An overall trend was observed as the higher flexibility in the TNR structure leads to lower capillary flow. After testing samples derived from relevant cells harboring expanded TNR sequences, it is concluded that this approach may transform into a reagent-free and pump-free biosensing platform to detect microsatellite expansion diseases.