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WO2006129845A1 - Appareil et procede pour fabriquer une nanoconstruction - Google Patents

Appareil et procede pour fabriquer une nanoconstruction Download PDF

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Publication number
WO2006129845A1
WO2006129845A1 PCT/JP2006/311237 JP2006311237W WO2006129845A1 WO 2006129845 A1 WO2006129845 A1 WO 2006129845A1 JP 2006311237 W JP2006311237 W JP 2006311237W WO 2006129845 A1 WO2006129845 A1 WO 2006129845A1
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WIPO (PCT)
Prior art keywords
nanostructure
dna
microchannel
temperature
self
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Application number
PCT/JP2006/311237
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English (en)
Japanese (ja)
Inventor
Satoshi Murata
Kotaro Somei
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Tokyo Institute Of Technology
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Tokyo Institute Of Technology filed Critical Tokyo Institute Of Technology
Publication of WO2006129845A1 publication Critical patent/WO2006129845A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA

Definitions

  • the present invention relates to a nanostructure production apparatus and method suitable for use in producing a nanostructure by self-assembling components such as DNA tiles, for example.
  • This application claims priority on the basis of Japanese Patent Application No. 2005-164203 filed on June 3, 2005 in Japan, and this application is incorporated herein by reference. .
  • DNA molecules have attracted attention as components of nanostructures.
  • DNA tiles, DNA junctions, DNA polygons, and DNA polyhedra have been actively studied.
  • Each of these DNA molecules has a different shape and number and position of sticky ends (single-stranded parts that do not form a double helix structure), and the structures formed by self-assembly are also different.
  • the DNA tile is expected as a component that can be used to create various nanostructures because the specific binding of each DNA tile can be designed freely by designing the four sticky ends circled in Fig. 10. Yes.
  • the self-assembly process of DNA tiles corresponds mathematically to a one-dimensional cellular automaton model called a Wang tile, and it is possible to perform some kind of parallel computation using this algorithmic self-assembly (Algorithmic Self Assembly).
  • Algorithmic Self Assembly -assembly
  • a DNA tile set with a sticky end corresponding to an XOR (exclusive OR) operation is used as a state transition rule, a two-dimensional crystal (nanostructure) of a DNA tile having a shellpin skifractal structure is obtained.
  • XOR exclusive OR
  • stepwise self-assembly in which two DNA tiles are placed in a test tube to advance self-assembly hierarchically, has been studied (for example, non-patent literature). 2).
  • FIG. 11A by adding DNA tiles with index numbers 1 to 16 in a hierarchical manner, a fully-addressable nanostructure with 4 ⁇ 4 vertical and horizontal DNA tiles as shown in FIG. 11B can be obtained. It can be realized.
  • FIG. 11B each DNA tile is conceptually shown as a square.
  • self-assembly can be performed relatively reliably.
  • the experimental operation is complicated, and it is difficult to control environmental parameters such as DNA tile concentration and temperature, as described above. There was a problem that nanostructures could not be produced.
  • An object of the present invention is to provide a device and a method for producing an effective nanostructure.
  • a nanostructure fabrication apparatus is a nanostructure fabrication apparatus that creates a nanostructure in a microchannel, and the microchannel includes a plurality of microchannels.
  • a method for manufacturing a nanostructure according to the present invention is a method for manufacturing a nanostructure in a microchannel, and the microchannel includes: Divided into a plurality of regions, a solution containing self-assembled components of the nanostructure is supplied to at least a portion of the microchannel, and the solution is provided downstream of the microchannel.
  • the nanostructure is produced in a reaction vessel in which a seed crystal of the nanostructure is fixed.
  • FIG. 1 is a diagram showing an external configuration of a microfluidic device in the present embodiment.
  • FIG. 2 is a diagram showing the shape of a microchannel.
  • FIG. 3 is a diagram showing the relationship between the position in the microchannel and the capillary force.
  • FIG. 4 is a diagram showing the flow of the solution when the solution is dropped into the service port of the microchannel.
  • FIG. 5 is a diagram showing a temperature profile of the microfluidic device when a voltage of 24 V is applied to the heater.
  • FIG. 6A and FIG. 6B are diagrams showing a conventional stepwise self-assembly using a one-pot reaction and a stepwise self-assembly using a microfluidic device.
  • FIG. 7 is a diagram showing an example of a nanostructure produced when hierarchical algorithmic self-assembly is performed using a microfluidic device.
  • FIG. 8 is a diagram showing an example of a microchannel divided into a plurality of regions.
  • FIG. 9 is a diagram showing another example of a microchannel divided into a plurality of regions.
  • FIG. 10 is a diagram schematically showing an example of a DNA tile.
  • FIG. 11A and FIG. 11B are diagrams for explaining a method for producing a nanostructure by a conventional stepwise self-assembly.
  • the present invention is applied to a microfluidic device in which DNA tiles are self-assembled in a reaction field in a microchannel (diameter / zm to several hundreds of m) and a nanostructure is produced. It is applied.
  • the microfluidic device 1 in the present embodiment is mainly formed by laminating a silicon substrate 11 on a glass substrate 10.
  • a recess 12 is formed in the center of the silicon substrate 11, and the bottom surface of the recess 12 is a microchannel 13 to 13 for flowing a solution in which a DNA tile is dissolved (hereinafter, there is no need to distinguish between them).
  • Microchannel 13 is a microchannel 13 to 13 for flowing a solution in which a DNA tile is dissolved
  • the number of microchannels 13 is three, but it is needless to say that the number is not limited to this number.
  • the microchannel 13 has an open surface so that a nanostructure crystal during or after fabrication can be observed with an AFM (Atomic Force Microscope).
  • AFM Anamic Force Microscope
  • the recess 12 is filled with oil such as mineral oil used in PCR (Polymerase Chain Reaction).
  • a heater 14 made of ITO (Indium Tin Oxide) and a temperature sensor 15 to 15 hereinafter, it is necessary to distinguish between the glass substrate 10 and the silicon substrate 11). If there is no
  • the microchannel 13 includes a service port 20 for dropping a solution in which a DNA tile is dissolved, and a service port 20 described later.
  • a stop valve 21 having a function to perform a reaction
  • a reaction chamber 22 that is a reaction field in which DNA tiles self-assemble
  • a mechanical pump 23 are also configured.
  • the reaction chamber 22 is fixed with an initial strand that becomes a seed crystal of the nanostructure.
  • the microchannel 13 is designed so that the flow path width gradually becomes narrower from the end of the service port 20 to the first pump 23, and as a result, the capillary force is increased.
  • the direction of the service port 20 is expressed as upstream
  • the direction of the mechanical pump 23 is expressed as downstream.
  • FIG. 3 shows the relationship between the position in the microchannel 13 and the capillary force.
  • the flow path width becomes narrower as it goes downstream, and the capillary force also becomes stronger accordingly.
  • the stop valve 21 the capillary force suddenly increases as the flow path width suddenly narrows.
  • the capillary force decreases slightly as the flow path width increases slightly. ing.
  • the flow path width is narrowed again to the same width as the stop valve 21, and accordingly, the capillary force becomes as strong as the position of the stop valve 21.
  • a nanostructure crystal is grown around the seed crystal.
  • unnecessary by-products around the seed crystal are washed away, and a large nanostructure can be produced.
  • crystal growth can be suppressed by growing the crystal along the flow of the solution.
  • conventional one-pot reactions such as dropping ligase into the service port 20 to connect DNA tiles, or dropping restriction enzymes to cleave specific sites, etc. Difficult reactions can be handled freely.
  • FIG. 5 shows the temperature profile of the microfluidic device 1 when a voltage of 24 V is applied to the heater 14. As shown in FIG. 5, a uniform temperature field can be obtained in the three microchannels 13-13.
  • the highest temperature at which DNA tiles can be bonded to each other is preferable. This is because the bonding force between the non-complementary sticky ends is weaker than that between the complementary sticky ends, and thus defects can be suppressed by increasing the temperature as much as possible.
  • microfluidic device 1 it is also possible to extend this and perform algorithmic self-assembly using the microfluidic device 1 in a hierarchical manner.
  • the complex as shown in FIG. Patterned nanostructures can be fabricated.
  • the sequences of the sticky ends can be made the same in each DNA tile set, it is not necessary to increase the number of sticky end sequences.
  • each protein has a unique function, it is necessary to bind it to a nanostructure that also has DNA tiling power, so that a system with a useful function, for example, a group of enzymes necessary to degrade a specific molecule
  • a DNA lattice or the like in which (protein group) is optimally arranged can be constructed.
  • the solution conditions metal ion concentration and other DN
  • Various DNA effector elements whose shapes change depending on the presence of the A sequence may be dropped on the service port 20.
  • a DNA actor element between DNA tiles, for example, it is possible to construct a filter made of a lattice (the size of the mesh is dynamically changed) that recognizes and passes only specific molecules.
  • a metal colloid (for example, gold colloid) nanoparticle bonded with a DNA binding linker may be dropped onto the service port 20. By arranging them in a row, it becomes a nanowire and a wiring pattern can be formed.
  • microfluidic device 1 various chemically synthesized nanoelectronic devices (electronic elements having rectification and amplification effects) bonded with a DNA binding linker are dropped onto the service port 20.
  • a DNA binding linker bonded with a DNA binding linker
  • each microchannel 13 has been described as one.
  • the present invention is not limited to such a configuration, and the microchannel is divided into a plurality of areas, and each area is divided into a plurality of areas.
  • a service port may be provided.
  • DNA tile formation from multiple single-stranded DNAs ⁇
  • Substructure A formation from two DNA tiles ⁇
  • Substructure B formation from two substructures A Each target nanostructure Steps can be performed in each of the areas described above.
  • Each region is controlled to an optimum temperature. In general, it is preferable to increase the initial stage of fabrication of nanostructures to a high temperature and to a low temperature as the stage progresses.
  • FIG. 8 An example of such a microchannel is shown in FIG.
  • the microchannel 30 shown in FIG. 8 is divided into first to fourth regions 31 to 31, and the first region 31 is heated to a high temperature and the second to third regions 31 to 31 are divided.
  • the 1 4 1 regions 31 and 31 are controlled at a medium temperature, and the fourth region 31 is controlled at a low temperature.
  • Service port 3
  • a solution in which single-stranded DNA is dissolved is dropped into 2 and 32, and the service port 32 has a DN.
  • the solution in which the A tile is dissolved is dropped, and the service port 32 has a sub-structure of the DNA tile.
  • a DNA tile is formed from a plurality of single-stranded DNAs in the second region 31, and two in the third region 31.
  • FIG. 9 Another example of such a microchannel is shown in FIG.
  • the microchannel 40 shown in FIG. 9 is divided into first to ninth regions 41 to 41, and the first, third, and sixth regions 41, 41,
  • the ninth region 41 is controlled to a low temperature. Also, service ports 42-42 have DN
  • the solution in which the tile is dissolved is dropped, and the solution in which the DNA tile is dissolved is dropped in the service port 42.
  • the first, third, and sixth regions 41, 41, and 41 are completely decomposed into single-stranded DNA.
  • the second, fourth, and seventh areas are completely decomposed into single-stranded DNA.
  • DNA tiles are formed from a plurality of single-stranded DNAs, and the fifth and eighth regions are formed.
  • the DNA tile substructure is formed from the two DNA tiles, and the ninth region
  • region 41 two DNA tile substructures and nanostructures are formed.
  • the DNA tile itself can also be fabricated in the microchannel, thereby making it possible to fabricate nanostructures with a lower defect rate.
  • the service port includes not only single-stranded DNA and DNA tile, but also enzyme groups such as ligase and restriction enzyme, protein, DNA activator element and various nanoparticles, It ’s okay to drop nanodevices.
  • motifs such as DNA junctions, DNA polygons, DNA polyhedra, etc., which are explained using DNA tiles as examples of DNA molecules that are constituent elements of nanostructures, are used. You may use. If the component is not limited to single-stranded DNA or DNA molecules but can be self-assembled, single-stranded RN A or other components such as RNA molecules or proteins may be used.

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Abstract

Selon l'invention, on verse goutte à goutte une solution dans un orifice de maintenance (20) dans un microcanal (13) puis elle est acheminée à une pompe capillaire (23) grâce à la force capillaire. La température dans une chambre de réaction (22), dans laquelle un brin de démarrage servant de germe de cristal pour une nanoconstruction est fixé, est régulée avec un dispositif de chauffage et un capteur de température. Lors de la fabrication de la nanoconstruction, par exemple, les solutions contenant chacune une seule tuile d'ADN qui y est dissoute sont successivement versées goutte à goutte dans l'orifice de maintenance (20) et les cristaux de la nanoconstruction croissent autour du germe de cristal dans les écoulements de solution.
PCT/JP2006/311237 2005-06-03 2006-06-05 Appareil et procede pour fabriquer une nanoconstruction WO2006129845A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2005164203A JP2006334741A (ja) 2005-06-03 2005-06-03 ナノ構造物の作製装置及び方法
JP2005-164203 2005-06-03

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EP2230504B1 (fr) * 2008-01-08 2013-04-24 Nippon Telegraph and Telephone Corporation Unité de pompage capillaire et cuve à circulation

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2003522963A (ja) * 2000-02-18 2003-07-29 アクララ バイオサイエンシーズ， インコーポレイテッド 複数部位反応デバイスおよび方法
US20040091399A1 (en) * 2002-11-11 2004-05-13 Chung Kwang Hyo Device for controlling fluid using surface tension
WO2004050967A1 (fr) * 2002-12-03 2004-06-17 Motorola Inc Liaison de carbone nanomorphe a un acide nucleique
JP2004194652A (ja) * 2002-12-06 2004-07-15 Dainippon Ink & Chem Inc 溶解性物質付着流路を有するマイクロ流体素子及びその使用方法
JP2005091246A (ja) * 2003-09-19 2005-04-07 Fujitsu Ltd デバイスおよびその製造方法

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2003522963A (ja) * 2000-02-18 2003-07-29 アクララ バイオサイエンシーズ， インコーポレイテッド 複数部位反応デバイスおよび方法
US20040091399A1 (en) * 2002-11-11 2004-05-13 Chung Kwang Hyo Device for controlling fluid using surface tension
WO2004050967A1 (fr) * 2002-12-03 2004-06-17 Motorola Inc Liaison de carbone nanomorphe a un acide nucleique
JP2004194652A (ja) * 2002-12-06 2004-07-15 Dainippon Ink & Chem Inc 溶解性物質付着流路を有するマイクロ流体素子及びその使用方法
JP2005091246A (ja) * 2003-09-19 2005-04-07 Fujitsu Ltd デバイスおよびその製造方法

Non-Patent Citations (2)

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Title
JUNCKER D. ET AL.: "Autonomous Microfluidic Capillary System", ANALYTICAL CHEMISTRY, AMERICAN CHEMICAL SOCIETY, vol. 74, no. 24, 15 December 2002 (2002-12-15), pages 6139 - 6144, XP001162255 *
WINFREE E. ET AL.: "Tile Sets: Error Correction for Algorithmic Self-Assembly", LECTURE NOTES IN COMPUTER SCIENCE (DNA COMPUTING), DE, SPRINGER, vol. 2943, 2004, pages 126 - 144, XP019002510 *

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