title: 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.
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 constant KsTTTATA-TATAAA= ka/kd≈ 106M-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.
authors.affiliation: Institute of Physical Chemistry, Polish Academy of Sciences , Kasprzaka 44/52, 01-224 Warsaw, Poland.
authors.forename: Katarzyna
authors.identifier: http://orcid.org/0000-0002-3526-9208
authors.initials: K
authors.lastname: Bartold
authors.affiliation: Institute of Physical Chemistry, Polish Academy of Sciences , Kasprzaka 44/52, 01-224 Warsaw, Poland.
authors.forename: Agnieszka
authors.identifier: http://orcid.org/0000-0003-2377-5493
authors.initials: A
authors.lastname: Pietrzyk-Le
authors.affiliation: Institute of Physical Chemistry, Polish Academy of Sciences , Kasprzaka 44/52, 01-224 Warsaw, Poland.|Department of Chemistry, University of North Texas , 1155 Union Circle, No. 305070, Denton, Texas 76203-5017, United States.
authors.forename: Tan-Phat
authors.initials: TP
authors.lastname: Huynh
authors.affiliation: Institute of Physical Chemistry, Polish Academy of Sciences , Kasprzaka 44/52, 01-224 Warsaw, Poland.
authors.forename: Zofia
authors.initials: Z
authors.lastname: Iskierko
authors.affiliation: Institute of Physical Chemistry, Polish Academy of Sciences , Kasprzaka 44/52, 01-224 Warsaw, Poland.|Department of Chemistry, University of North Texas , 1155 Union Circle, No. 305070, Denton, Texas 76203-5017, United States.
authors.forename: Marta
authors.initials: M
authors.lastname: Sosnowska
authors.affiliation: Institute of Physical Chemistry, Polish Academy of Sciences , Kasprzaka 44/52, 01-224 Warsaw, Poland.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Noworyta
authors.affiliation: Institute of Physical Chemistry, Polish Academy of Sciences , Kasprzaka 44/52, 01-224 Warsaw, Poland.
authors.forename: Wojciech
authors.initials: W
authors.lastname: Lisowski
authors.affiliation: Department of Chemistry, University of Milan , Via Golgi 19, I-20133 Milan, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Sannicolò
authors.affiliation: Department of Chemistry, University of Milan , Via Golgi 19, I-20133 Milan, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Cauteruccio
authors.affiliation: Department of Chemistry, University of Milan , Via Golgi 19, I-20133 Milan, Italy.
authors.forename: Emanuela
authors.initials: E
authors.lastname: Licandro
authors.affiliation: Department of Chemistry, University of North Texas , 1155 Union Circle, No. 305070, Denton, Texas 76203-5017, United States.
authors.forename: Francis
authors.initials: F
authors.lastname: D'Souza
authors.affiliation: Institute of Physical Chemistry, Polish Academy of Sciences , Kasprzaka 44/52, 01-224 Warsaw, Poland.|Faculty of Mathematics and Natural Sciences, School of Sciences, Cardinal Stefan Wyszynski University , Woycickiego 1/3, 01-938 Warsaw, Poland.
authors.forename: Wlodzimierz
authors.identifier: http://orcid.org/0000-0003-3586-5170
authors.initials: W
authors.lastname: Kutner
keywords: DNA/RNA analogue; bis(2,2′-bithien-5-yl)methane monomer; capacitive impedimetry; chemosensor; molecularly imprinted polymer; oligonucleotide; peptide nucleic acid; piezoelectric microgravimetry