title: A sol-gel-based microfluidics system enhances the efficiency of RNA aptamer selection.
abstract: RNA and DNA aptamers that bind to target molecules with high specificity and affinity have been a focus of diagnostics and therapeutic research. These aptamers are obtained by SELEX often requiring many rounds of selection and amplification. Recently, we have shown the efficient binding and elution of RNA aptamers against target proteins using a microfluidic chip that incorporates 5 sol-gel binding droplets within which specific target proteins are imbedded. Here, we demonstrate that our microfluidic chip in a SELEX experiment greatly improved selection efficiency of RNA aptamers to TATA-binding protein, reducing the number of selection cycles needed to produce high affinity aptamers by about 80%. Many aptamers were identical or homologous to those isolated previously by conventional filter-binding SELEX. The microfluidic chip SELEX is readily scalable using a sol-gel microarray-based target multiplexing. Additionally, we show that sol-gel embedded protein arrays can be used as a high-throughput assay for quantifying binding affinities of aptamers.
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University , Ithaca, New York.
authors.forename: Ji-Young
authors.initials: JY
authors.lastname: Ahn
authors.forename: Minjoung
authors.initials: M
authors.lastname: Jo
authors.forename: Pooja
authors.initials: P
authors.lastname: Dua
authors.forename: Dong-Ki
authors.initials: DK
authors.lastname: Lee
authors.forename: Soyoun
authors.initials: S
authors.lastname: Kim
mesh_terms: D052157:Aptamers, Nucleotide / Q000032:analysis / Q000737:chemistry* / Q000235:genetics*; D001483:Base Sequence; D055767:Immobilized Proteins / Q000032:analysis / Q000737:chemistry; D044085:Microfluidics / Q000295:instrumentation*; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D036103:Nanotechnology / Q000295:instrumentation; D052156:SELEX Aptamer Technique / Q000295:instrumentation*; D012680:Sensitivity and Specificity; D035181:TATA-Box Binding Protein / Q000032:analysis / Q000737:chemistry*
chemical_list: D052157:Aptamers, Nucleotide; D055767:Immobilized Proteins; D035181:TATA-Box Binding Protein