Programmed Transfer of Sequence Information into a Molecularly Imprinted Polymer for Hexakis(2,2 '-bithien-5-yl) DNA Analogue Formation toward Single-Nucleotide-Polymorphism Detection

A1 Journal article (refereed)


Internal Authors/Editors


Publication Details

List of Authors: Katarzyna Bartold, Agnieszka Pietrzyk-Le, Tan-Phat Huynh, Zofia Iskierko, Marta Sosnowska, Krzysztof Noworyta, Wojciech Lisowski, Francesco Sannicolò, Silvia Cauteruccio, Emanuela Licandro, Francis D’Souza, Wlodzimierz Kutner
Publisher: ACS
Publication year: 2017
Journal: ACS Applied Materials and Interfaces
Journal acronym: ACS APPL MATER INTER
Volume number: 9
Issue number: 4
Start page: 3948
End page: 3958
Number of pages: 11
ISSN: 1944-8244
eISSN: 1944-8252


Abstract

A new strategy of simple, inexpensive, rapid, and label free single-nucleotide-polymorphism (SNP) detection using robust chemosensors with piezomicrogravimetric, surface plasmon resonance, or capacitive impedimetry (CI) signal transduction is reported. Using these chemosensors, selective detection of a genetically relevant oligonucleotide under FIA conditions within 2 min is accomplished. An invulnerable-to-nonspecific interaction molecularly imprinted polymer (MIP) with electrochemically synthesized probes of hexameric 2,2'-bithien-5-yl DNA analogues discriminating single purine nucleobase mismatch at room temperature was used. With density functional theory modeling, the synthetic procedures developed, and isothermal titration calorimetry quantification, adenine (A)- or thymine (T)-substituted 2,2'-bithien-5-yl functional monomers capable of Watson Crick nucleobase pairing with the TATAAA oligodeoxyribonucleotide template or its peptide nucleic acid (PNA) analogue were designed. Characterized by spectroscopic techniques, molecular cavities exposed the ordered nucleobases on the 2,2'-bithien-5-yl polymeric backbone of the TTTATA hexamer probe designed to hybridize the complementary TATAAA template. In that way, an artificial TATAAA-promoter sequence was formed in the MIP. The purine nucleobases of this sequence are known to be recognized by RNA polymerase to initiate the transcription in eukaryotes. The hexamer strongly hybridized TATAAA with the complex stability TTTATA-TATAAA constant K-s(TTTATA-TATAAA)approximate to k(a)/k(d) 10(6) M-1, as high as that characteristic for longer-chain DNA PNA hybrids. The CI chemosensor revealed a 5 nM limit of detection, quite appreciable as for the hexadeoxyribonucleotide. Molecular imprinting increased the chemosensor sensitivity to the TATAAA analyte by over 4 times compared to that of the nonimprinted polymer. The herein-devised detection platform enabled the generation of a library of hexamer probes for typing the majority of SNP probes as well as studying a molecular mechanism of the complex transcription machinery, physics of single polymer molecules, and stable genetic nanomaterials.


Keywords

bis(2,2 '-bithien-5-yl)methane monomer, Capacitive impedimetry, chemosensor, DNA/RNA analogue, molecularly imprinted polymer, peptide nucleic acid, Piezoelectric microgravimetry

Last updated on 2019-18-01 at 04:43