title: Temperature-controlled microintaglio printing for high-resolution micropatterning of RNA molecules.
abstract: We have developed an advanced microintaglio printing method for fabricating fine and high-density micropatterns and applied it to the microarraying of RNA molecules. The microintaglio printing of RNA reported here is based on the hybridization of RNA with immobilized complementary DNA probes. The hybridization was controlled by switching the RNA conformation via the temperature, and an RNA microarray with a diameter of 1.5 µm and a density of 40,000 spots/mm(2) with high contrast was successfully fabricated. Specifically, no size effects were observed in the uniformity of patterned signals over a range of microarray feature sizes spanning one order of magnitude. Additionally, we have developed a microintaglio printing method for transcribed RNA microarrays on demand using DNA-immobilized magnetic beads. The beads were arrayed on wells fabricated on a printing mold and the wells were filled with in vitro transcription reagent and sealed with a DNA-immobilized glass substrate. Subsequently, RNA was in situ synthesized using the bead-immobilized DNA as a template and printed onto the substrate via hybridization. Since the microintaglio printing of RNA using DNA-immobilized beads enables the fabrication of a microarray of spots composed of multiple RNA sequences, it will be possible to screen or analyze RNA functions using an RNA microarray fabricated by temperature-controlled microintaglio printing (TC-µIP).
authors.affiliation: Department of Bioengineering, School of Engineering, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-8656, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Kobayashi
authors.affiliation: Department of Bioengineering, School of Engineering, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-8656, Japan; Japan Science and Technology Agency, CREST, Chiyoda, Tokyo 102-0075, Japan; Green Devices Research Center, School of Materials Science, Japan Advanced Institute of Science and Technology, 1-1 Asahidai, Nomi, Ishikawa 923-1292, Japan. Electronic address: biyani@jaist.ac.jp.
authors.forename: Manish
authors.initials: M
authors.lastname: Biyani
authors.affiliation: Department of Bioengineering, School of Engineering, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-8656, Japan; Japan Science and Technology Agency, CREST, Chiyoda, Tokyo 102-0075, Japan.
authors.forename: Shingo
authors.initials: S
authors.lastname: Ueno
authors.affiliation: Department of Bioengineering, School of Engineering, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-8656, Japan.
authors.forename: Subhashini Raj
authors.initials: SR
authors.lastname: Kumal
authors.affiliation: Department of Bioengineering, School of Engineering, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-8656, Japan.
authors.forename: Hiromi
authors.initials: H
authors.lastname: Kuramochi
authors.affiliation: Department of Bioengineering, School of Engineering, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-8656, Japan; Japan Science and Technology Agency, CREST, Chiyoda, Tokyo 102-0075, Japan. Electronic address: ichiki@bioeng.t.u-tokyo.ac.jp.
authors.forename: Takanori
authors.initials: T
authors.lastname: Ichiki
mesh_terms: D001483:Base Sequence; D015374:Biosensing Techniques; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D011327:Printing; D012313:RNA / Q000737:chemistry* / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D012313:RNA
keywords: High-throughput screening; Intaglio printing; Microarray; Parallel spotting; RNA patterning; in situ synthesis