この作品は、米海軍研究若手研究プログラム賞N000141110914、海軍研究助成N000141010827のオフィス、NSFキャリアアワードCCF1054898、NIHディレクターの新イノベーター賞1DP2OD007292のオフィスと（PY）は生物学的にインスパイア工学部スタートアップ基金ウィス研究所によって資金を供給させました清華 - 北京（BW）はライフサイエンススタートアップ基金センター。

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The authors declare competing financial interests.

構造形成工程では、自己組織化DNAナノ構造のDNA鎖の混合物（ 例えば 、15 mM）のマグネシウムカチオンの適切な濃度を維持することが重要です。同様に、アガロースゲル特性評価/精製工程では、ゲル電気泳動中にDNAナノ構造を保持するゲルランニング緩衝液中で、適切なマグネシウムカチオン濃度（ 例えば 、10ミリモル）を維持することが重要です。 28T矩形構造×24Hのために、?...

5,8,10 - - 13,17,23またはRNA 7,22定期的な3,4,7、前の核酸の自己組立作業1-25は、DNA 2を含む複雑な構造の様々な成功構築につながっています22とアルゴリズム5二次元格子、リボン10,12およびチューブ4,12,13、3次元結晶17、多面体11と有限、2Dは7,8を整形します。 21,25 - 16,18 -特に効果的な方法は、単一の足場鎖が複雑な形状9,14を形成するために、多くの短い補助ステープルストランドによって折り畳まれることにより足場のDNA折り紙、です。 我々は最近、一本鎖のタイル（SST）を使用して、所定の2次元形状を有する個別のナノ構造を構築するための方法を報告し、DNA折り紙26に匹敵する複雑さを有する構造を示しました。このarticleは私達の初期の作品26の適応であり、正確に所定の寸 ​​法（幅と長さ）および形態と洗練された有限の2次元形状に個別にアドレス指定するSSTを配置するための詳細なプロトコルを記述します。 SST方式の一つの重要な利点は、そのモジュール性です。構造体のすべてのコンポーネントのSSTアセンブリでモジュール構造単位として機能し、これらの海面水温の異なるサブセットは、異なる形状を作り出します。したがって、我々は短い、合成DNA鎖から所定の大きさや形状でナノ構造を構築するための一般的なプラットフォームを確立しました。 海面水温は、それぞれ10または11ヌクレオチド長の4つのドメイン（ 図1A）を含みます。海面水温は、その平行ヘリックスは、クロスオーバー結合によって一緒に保持されたDNA格子を作成するように結合します。各クロスオーバーは、リン酸が見やすくするための図で人為的に延伸されるドメイン2と3の間にリン酸塩です。クロスオーバーは、（離れて2ヘリカルターン（21塩基）を離間されています&lt;強い&gt;図1B）。複合長方形はヘリックスとヘリカル巻き数にそれらの寸法によって参照されます。たとえば、6ヘリックス幅で、8ヘリカルが長くなります長方形が8Tの長方形×6Hとして参照されます。海面水温は、取り残さ追加、またはその他の任意の形状と大きさ（ 図1C）の構造を作成するために再配置することができます。例えば、長方形のデザインは、所望の長さと半径（ 図1D）で管にロールバックすることができます。 あるいは、矩形SST格子はそれぞれ3 NMが7nmでによって、SSTの画素からなる分子キャンバスとみなすことができます。本研究では、310全長内部海面水温、左右の境界を構成する24の完全長海面水温、および上部と下部の境界を形成する28の半分の長さの海面水温の分子キャンバスを使用しています。キャンバスは、クロスオーバーによってリンクされた24の二重らせんを有し、各螺旋は28ヘリカルターン（294塩基）を含有し、したがってと呼ばれます28T矩形キャンバス×24H。 28Tキャンバス×24Hは、M13ファージの足場から作成されたDNA折り紙構造と同様の分子量を有します。

<table><tbody><tr><td>DNA Strands&amp;nbsp;</td><td>Integrated DNA Technology</td><td></td><td>Section 3.1</td></tr><tr><td>SYBR Safe DNA gel stain</td><td>Invitrogen</td><td>S33102</td><td>Section 3.4.2</td></tr><tr><td>Freeze&#039;N Squeeze DNA Gel Extraction Spin Columns</td><td>BIO-RAD</td><td>731-6166</td><td>Section 3.6</td></tr><tr><td>Bruker&#039;s Sharp Nitride Lever Probes</td><td>Bruker AFM Probes</td><td>SNL10</td><td>Section 4.3</td></tr><tr><td>Safe Imager 2.0 Blue Light Transilluminator</td><td>Invitrogen</td><td>G6600</td><td>Section 3.6</td></tr><tr><td>Centrifuge 5430R</td><td>Eppendorf</td><td>5428 000.414</td><td>Section 3.6</td></tr><tr><td>Transmission Electron Microscope&amp;nbsp;</td><td>Jeol</td><td>Jem 1400</td><td>Section 7.4</td></tr><tr><td>Multimode 8</td><td>Veeco</td><td></td><td>Section 4</td></tr><tr><td>Typhoon FLA 9000 Laser Scanner</td><td>GE Heathcare Life Sciences</td><td>28-9558-08</td><td>Section 3.5</td></tr><tr><td>Ultrapure Distilled water</td><td>Invitrogen</td><td>10977-023</td><td>Section 3.7.1</td></tr><tr><td>Mica disk</td><td>SPI Supplies</td><td>12001-26-2</td><td>Section 4.1</td></tr><tr><td>Steel mounting disk</td><td>Ted Pella, Inc.</td><td>16218</td><td>Section 4.1</td></tr><tr><td>carbon coated copper grid for TEM</td><td>Electron Microscopy Sciences</td><td>FCF400-Cu</td><td>Section 7.2</td></tr><tr><td>tweezers</td><td>Dumont</td><td>0203-N5AC-PO</td><td>Section 7.31</td></tr><tr><td>glow discharge system</td><td>Quorum Technologies</td><td>K100X</td><td>Section 7.2</td></tr><tr><td>DNA Engine Tetrad 2 Peltier Thermal Cycler</td><td>BIO-RAD</td><td>PTC&amp;ndash;0240G</td><td>Section 3.3</td></tr><tr><td>Owl Easycast B2 Mini Gel Electrophoresis Systems</td><td>ThermoScientific</td><td>B2</td><td>Section 3.4.3</td></tr><tr><td>Seekam LE Agarose 500G</td><td>Lonza</td><td>50004</td><td>Section 3.4.1</td></tr><tr><td>GeneRuler 1kb Plus DNA Ladder, Ready-To-Use 75-20000bp</td><td>ThermoScientific</td><td>SM1333</td><td>Section 3.4.4</td></tr><tr><td>Nanodrop 2000c UV-vis Spectrophotometer</td><td>ThermoScientific</td><td></td><td>Section 3.7</td></tr><tr><td>0.2 um filter</td><td>Corning Inc.</td><td>431219</td><td>Section 7.1.2</td></tr></tbody></table>

self-assembly of complex two-dimensional shapes from single-stranded dna tiles

Current methods in DNA nano-architecture have successfully engineered a variety of 2D and 3D structures using principles of self-assembly. In this article, we describe detailed protocols on how to fabricate sophisticated 2D shapes through the self-assembly of uniquely addressable single-stranded DNA tiles which act as molecular pixels on a molecular canvas. Each single-stranded tile (SST) is a 42-nucleotide DNA strand composed of four concatenated modular domains which bind to four neighbors during self-assembly. The molecular canvas is a rectangle structure self-assembled from SSTs. A prescribed complex 2D shape is formed by selecting the constituent molecular pixels (SSTs) from a 310-pixel molecular canvas and then subjecting the corresponding strands to one-pot annealing. Due to the modular nature of the SST approach we demonstrate the scalability, versatility and robustness of this method. Compared with alternative methods, the SST method enables a wider selection of information polymers and sequences through the use of de novo designed and synthesized short DNA strands.

海面水温の自己集合（図1）は、図2に示すように、28T矩形×24Hをもたらす。別の海面水温のためのDNA配列は、ストレプトアビジン標識（図3及び4）の形質転換を可能にするように最適化/改変することができます様々なサイズのチューブや長方形を形成するためにチューブに長方形（ 図5）、海面水温のプログラム可能な自己集合（図10）?...

Watch this Scientific Journal Video about Self-assembly of Complex Two-dimensional Shapes from Single-stranded DNA Tiles at JoVE.com

Self-assembly of Complex Two-dimensional Shapes from Single-stranded DNA Tiles

DNA tiling is an effective approach to make programmable nanostructures. We describe the protocols to construct complex two-dimensional shapes by the self-assembly of single-stranded DNA tiles.

一本鎖DNAタイルから複雑な2次元形状の自己集合

Current methods in DNA nano-architecture have successfully engineered a variety of 2D and 3D structures using principles of self-assembly. In this ...

self-assembly-complex-two-dimensional-shapes-from-single-stranded-dna

Universidad San Sebastian

Research

JoVE Journal

Chemistry

11.6K ビュー.  Tsinghua University. DNA tiling is an effective approach to make programmable nanostructures. We describe the protocols to construct complex two-dimensional shapes by the self-assembly of single-stranded DNA tiles.

ビデオ: Self-assembly of Complex Two-dimensional Shapes from Single-stranded DNA Tiles

Origami Inspired Self-assembly of Patterned and Reconfigurable Particles

There are numerous techniques such as photolithography, electron-beam lithography and soft-lithography that can be used to precisely pattern two dimensional (2D) structures. These technologies are mature, offer high precision and many of them can be implemented in a high-throughput manner. We leverage the advantages of planar lithography and combine them with self-folding methods1-20 wherein physical forces derived from surface tension or residual stress, are used to curve or fold planar structures into three dimensional (3D) structures. In doing so, we make it possible to mass produce precisely patterned static and reconfigurable particles that are challenging to synthesize.
In this paper, we detail visualized experimental protocols to create patterned particles, notably, (a) permanently bonded, hollow, polyhedra that self-assemble and self-seal due to the minimization of surface energy of liquefied hinges21-23 and (b) grippers that self-fold due to residual stress powered hinges24,25. The specific protocol described can be used to create particles with overall sizes ranging from the micrometer to the centimeter length scales. Further, arbitrary patterns can be defined on the surfaces of the particles of importance in colloidal science, electronics, optics and medicine. More generally, the concept of self-assembling mechanically rigid particles with self-sealing hinges is applicable, with some process modifications, to the creation of particles at even smaller, 100 nm length scales22, 26 and with a range of materials including metals21, semiconductors9 and polymers27. With respect to residual stress powered actuation of reconfigurable grasping devices, our specific protocol utilizes chromium hinges of relevance to devices with sizes ranging from 100 &mu;m to 2.5 mm. However, more generally, the concept of such tether-free residual stress powered actuation can be used with alternate high-stress materials such as heteroepitaxially deposited semiconductor films5,7 to possibly create even smaller nanoscale grasping devices.

We describe experimental details of the synthesis of patterned and reconfigurable particles from two dimensional (2D) precursors. This methodology can be used to create particles in a variety of shapes including polyhedra and grasping devices at length scales ranging from the micro to centimeter scale.

折り紙インスパイアパターン化され、再構成可能な粒子の自己組織化

There are numerous techniques such as photolithography, electron-beam lithography and soft-lithography that can be used to precisely pattern two ...

化学

Folding and Characterization of a Bio-responsive Robot from DNA Origami

The DNA nanorobot is a hollow hexagonal nanometric device, designed to open in response to specific stimuli and present cargo sequestered inside. Both stimuli and cargo can be tailored according to specific needs. Here we describe the DNA nanorobot fabrication protocol, with the use of the DNA origami technique. The procedure initiates by mixing short single-strand DNA staples into a stock mixture which is then added to a long, circular, single-strand DNA scaffold in presence of a folding buffer. A standard thermo cycler is programmed to gradually lower the mixing reaction temperature to facilitate the staples-to-scaffold annealing, which is the guiding force behind the folding of the nanorobot. Once the 60 hr folding reaction is complete, excess staples are discarded using a centrifugal filter, followed by visualization via agarose-gel electrophoresis (AGE). Finally, successful fabrication of the nanorobot is verified by transmission electron microscopy (TEM), with the use of uranyl-formate as negative stain.

DNA origami is a powerful method for fabricating precise nanoscale objects by programming the self-assembly of DNA molecules. Here we describe a protocol for the folding of a bio-responsive robot from DNA origami, its purification and negative staining for transmission electron microscopic imaging (TEM).

DNA折り紙からバイオ応答ロボットの折りたたみとキャラクタリゼーション

The DNA nanorobot is a hollow hexagonal nanometric device, designed to open in response to specific stimuli and present cargo sequestered inside. Both ...

Preparation of Mica and Silicon Substrates for DNA Origami Analysis and Experimentation

The designed nature and controlled, one-pot synthesis of DNA origami provides exciting opportunities in many fields, particularly nanoelectronics. Many of these applications require interaction with and adhesion of DNA nanostructures to a substrate. Due to its atomically flat and easily cleaned nature, mica has been the substrate of choice for DNA origami experiments. However, the practical applications of mica are relatively limited compared to those of semiconductor substrates. For this reason, a straightforward, stable, and repeatable process for DNA origami adhesion on derivatized silicon oxide is presented here. To promote the adhesion of DNA nanostructures to silicon oxide surface, a self-assembled monolayer of 3-aminopropyltriethoxysilane (APTES) is deposited from an aqueous solution that is compatible with many photoresists. The substrate must be cleaned of all organic and metal contaminants using Radio Corporation of America (RCA) cleaning processes and the native oxide layer must be etched to ensure a flat, functionalizable surface. Cleanrooms are equipped with facilities for silicon cleaning, however many components of DNA origami buffers and solutions are often not allowed in them due to contamination concerns. This manuscript describes the set-up and protocol for in-lab, small-scale silicon cleaning for researchers who do not have access to a cleanroom or would like to incorporate processes that could cause contamination of a cleanroom CMOS clean bench. Additionally, variables for regulating coverage are discussed and how to recognize and avoid common sample preparation problems is described.

Reproducible cleaning processes for substrates used in DNA origami research are described, including bench-top RCA cleaning and derivatization of silicon oxide. Protocols for surface preparation, DNA origami deposition, drying parameters, and simple experimental set-ups are illustrated.

DNA折り紙解析と実験のためのマイカとシリコン基板の作製

The designed nature and controlled, one-pot synthesis of DNA origami provides exciting opportunities in many fields, particularly nanoelectronics. Many of ...

Designing a Bio-responsive Robot from DNA Origami

Nucleic acids are astonishingly versatile. In addition to their natural role as storage medium for biological information1, they can be utilized in parallel computing2,3 , recognize and bind molecular or cellular targets4,5 , catalyze chemical reactions6,7 , and generate calculated responses in a biological system8,9. Importantly, nucleic acids can be programmed to self-assemble into 2D and 3D structures10-12, enabling the integration of all these remarkable features in a single robot linking the sensing of biological cues to a preset response in order to exert a desired effect.
Creating shapes from nucleic acids was first proposed by Seeman13, and several variations on this theme have since been realized using various techniques11,12,14,15 . However, the most significant is perhaps the one proposed by Rothemund, termed scaffolded DNA origami16. In this technique, the folding of a long (&gt;7,000 bases) single-stranded DNA 'scaffold' is directed to a desired shape by hundreds of short complementary strands termed 'staples'. Folding is carried out by temperature annealing ramp. This technique was successfully demonstrated in the creation of a diverse array of 2D shapes with remarkable precision and robustness. DNA origami was later extended to 3D as well17,18 .
 
The current paper will focus on the caDNAno 2.0 software19 developed by Douglas and colleagues. caDNAno is a robust, user-friendly CAD tool enabling the design of 2D and 3D DNA origami shapes with versatile features. The design process relies on a systematic and accurate abstraction scheme for DNA structures, making it relatively straightforward and efficient.
In this paper we demonstrate the design of a DNA origami nanorobot that has been recently described20. This robot is 'robotic' in the sense that it links sensing to actuation, in order to perform a task. We explain how various sensing schemes can be integrated into the structure, and how this can be relayed to a desired effect. Finally we use Cando21 to simulate the mechanical properties of the designed shape. The concept we discuss can be adapted to multiple tasks and settings.

DNA origami is a powerful method for fabricating precise nanoscale objects by programming the self-assembly of DNA molecules. Here, we describe how DNA origami can be utilized to design a robotic robot capable of sensing biological cues and responding by shape shifting, subsequently relayed to a desired effect.

DNA折り紙からバイオ応答ロボットの設計

Nucleic acids are astonishingly versatile. In addition to their natural role as storage medium for biological information1, they can be utilized in ...

生物工学

Stable DNA Motifs, 1D and 2D Nanostructures Constructed from Small Circular DNA Molecules

This article presents a detailed protocol for synthesis of small circular DNA molecules, annealing of circular DNA motifs, and construction of 1D and 2D DNA nanostructures. Over decades, the rapid development of DNA nanotechnology is attributed to the use of linear DNAs as the source materials. For example, the DAO (double crossover, antiparallel, odd half-turns) tile is well-known as a building block for construction of 2D DNA lattices; the core structure of DAO is made from two linear single-stranded (ss) oligonucleotides, like two ropes making a right hand granny&#160;knot. Herein, a new type of DNA tiles called cDAO (coupled DAO) are built using a small circular ss-DNA of c64nt or c84nt (circular 64 or 84 nucleotides) as the scaffold strand and several linear ss-DNAs as the staple strands. Perfect 1D and 2D nanostructures are assembled from cDAO tiles: infinite nanowires, nanospirals, nanotubes, nanoribbons; and finite nano-rectangles. Detailed protocols are described: 1) preparation by T4 ligase and purification by denaturing PAGE (polyacrylamide gel electrophoresis) of small circular oligonucleotides, 2) annealing of stable circular tiles, followed by native PAGE analysis, 3) assembling of infinite 1D nanowires, nanorings, nanospirals, infinite 2D lattices of nanotubes and nanoribbons, and finite 2D nano-rectangles, followed by AFM (Atomic Force Microscopy) imaging. The method is simple, robust, and affordable for most labs.

This article presents a detailed protocol for T4 ligation and denaturing PAGE purification of small circular DNA molecules, annealing and native PAGE analysis of circular tiles, assembling and AFM imaging of 1D and 2D DNA nanostructures, as well as agarose gel electrophoresis and centrifugation purification of finite DNA nanostructures.

ナノ構造構築小さな環状 DNA 分子から安定した DNA モチーフ、1 D および 2 D

This article presents a detailed protocol for synthesis of small circular DNA molecules, annealing of circular DNA motifs, and construction of 1D and 2D ...

DNA Origami-Mediated Substrate Nanopatterning of Inorganic Structures for Sensing Applications

Structural DNA nanotechnology provides a viable route for building from the bottom-up using DNA as construction material. The most common DNA nanofabrication technique is called DNA origami, and it allows high-throughput synthesis of accurate and highly versatile structures with nanometer-level precision. Here, it is shown how the spatial information of DNA origami can be transferred to metallic nanostructures by combining the bottom-up DNA origami with the conventionally used top-down lithography approaches. This allows fabrication of billions of tiny nanostructures in one step onto selected substrates. The method is demonstrated using bowtie DNA origami to create metallic bowtie-shaped antenna structures on silicon nitride or sapphire substrates. The method relies on the selective growth of a silicon oxide layer on top of the origami deposition substrate, thus resulting in a patterning mask for following lithographic steps. These nanostructure-equipped surfaces can be further used as molecular sensors (e.g., surface-enhanced Raman spectroscopy (SERS)) and in various other optical applications at the visible wavelength range owing to the small feature sizes (sub-10 nm). The technique can be extended to other materials through methodological modifications; therefore, the resulting optically active surfaces may find use in development of metamaterials and metasurfaces.

Here, we describe a protocol to create discrete and accurate inorganic nanostructures on substrates using DNA origami shapes as guiding templates. The method is demonstrated by creating plasmonic gold bowtie-shaped antennas on a transparent substrate (sapphire).

センシング用途のための無機構造のDNA折り紙媒介基板ナノパターニング

Structural DNA nanotechnology provides a viable route for building from the bottom-up using DNA as construction material. The most common DNA ...

Synthesis of Information-bearing Peptoids and their Sequence-directed Dynamic Covalent Self-assembly

This protocol presents the use of Lewis acidic multi-role reagents to circumvent kinetic trapping observed during the self-assembly of information-encoded oligomeric strands mediated by paired dynamic covalent interactions in a manner mimicking the thermal cycling commonly employed for the self-assembly of complementary nucleic acid sequences. Primary amine monomers bearing aldehyde and amine pendant moieties are functionalized with orthogonal protecting groups for use as dynamic covalent reactant pairs. Using a modified automated peptide synthesizer, the primary amine monomers are encoded into oligo(peptoid) strands through solid-phase submonomer synthesis. Upon purification by high-performance liquid chromatography (HPLC) and characterization by electrospray ionization mass spectrometry (ESI-MS), sequence-specific oligomers are subjected to high-loading of a Lewis acidic rare-earth metal triflate which both deprotects the aldehyde moieties and affects the reactant pair equilibrium such that strands completely dissociate. Subsequently, a fraction of the Lewis acid is extracted, enabling annealing of complementary sequence-specific strands to form information-encoded molecular ladders characterized by matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS). The simple procedure outlined in this report circumvents kinetic traps commonly experienced in the field of dynamic covalent assembly and serves as a platform for the future design of robust, complex architectures.

A protocol is presented for the synthesis of information-encoded peptoid oligomers and for the sequence-directed self-assembly of these peptoids into molecular ladders using amines and aldehydes as dynamic covalent reactant pairs and Lewis acidic rare-earth metal triflates as multi-role reagents.

情報を持つペプトイドの合成と、その配列指向動的共有自己集合

This protocol presents the use of Lewis acidic multi-role reagents to circumvent kinetic trapping observed during the self-assembly of information-encoded ...

Design and Synthesis of a Reconfigurable DNA Accordion Rack

DNA nanostructure-based mechanical systems or DNA nanomachines, which produce complex nanoscale motion in 2D and 3D in the nanometer to &#229;ngstr&#246;m resolution, show great potential in various fields of nanotechnology such as the molecular reactors, drug delivery, and nanoplasmonic systems. The reconfigurable DNA accordion rack, which can collectively manipulate a 2D or 3D nanoscale network of elements, in multiple stages in response to the DNA inputs, is described. The platform has potential to increase the number of elements that DNA nanomachines can control from a few elements to a network scale with multiple stages of reconfiguration.
In this protocol, we describe the entire experimental process of the reconfigurable DNA accordion rack of 6 by 6 meshes. The protocol includes a design rule and simulation procedure of the structures and a wet-lab experiment for synthesis and reconfiguration. In addition, analysis of the structure using TEM (transmission electron microscopy) and FRET (fluorescence resonance energy transfer) is included in the protocol. The novel design and simulation methods covered in this protocol will assist researchers to use the DNA accordion rack for further applications.

We describe the detailed protocol for design, simulation, wet-lab experiments, and analysis for a reconfigurable DNA accordion rack of 6 by 6 meshes.

再構成可能な DNA アコーディオン ラックの設計と合成

DNA nanostructure-based mechanical systems or DNA nanomachines, which produce complex nanoscale motion in 2D and 3D in the nanometer to &#229;ngstr&#246;m ...

Self-Assembly of Gamma-Modified Peptide Nucleic Acids into Complex Nanostructures in Organic Solvent Mixtures

Current strategies in DNA and RNA nanotechnology enable the self-assembly of a variety of nucleic acid nanostructures in aqueous or substantially hydrated media. In this article, we describe detailed protocols that enable the construction of nanofiber architectures in organic solvent mixtures through the self-assembly of uniquely addressable, single-stranded, gamma-modified peptide nucleic acid (&#947;PNA) tiles. Each single-stranded tile (SST) is a 12-base &#947;PNA oligomer composed of two concatenated modular domains of 6 bases each. Each domain can bind to a mutually complimentary domain present on neighboring strands using programmed complementarity to form nanofibers that can grow to microns in length. The SST motif is made of 9 total oligomers to enable the formation of 3-helix nanofibers. In contrast with analogous DNA nanostructures, which form diameter-monodisperse structures, these &#947;PNA systems form nanofibers that bundle along their widths during self-assembly in organic solvent mixtures. Self-assembly protocols described here therefore also include a conventional surfactant, Sodium Dodecyl Sulfate (SDS), to reduce bundling effects.

This article provides protocols for the design and self-assembly of nanostructures from gamma-modified peptide nucleic acid oligomers in organic solvent mixtures.

有機溶媒混合物中の複雑なナノ構造へのガンマ修飾ペプチド核酸の自己集合

Current strategies in DNA and RNA nanotechnology enable the self-assembly of a variety of nucleic acid nanostructures in aqueous or substantially hydrated ...

Simple, Affordable, and Modular Patterning of Cells using DNA

The relative positioning of cells is a key feature of the microenvironment that organizes cell-cell interactions. To study the interactions between cells of the same or different type, micropatterning techniques have proved useful. DNA Programmed Assembly of Cells (DPAC) is a micropatterning technique that targets the adhesion of cells to a substrate or other cells using DNA hybridization. The most basic operations in DPAC begin with decorating cell membranes with lipid-modified oligonucleotides, then flowing them over a substrate that has been patterned with complementary DNA sequences. Cells adhere selectively to the substrate only where they find a complementary DNA sequence. Non-adherent cells are washed away, revealing a pattern of adherent cells. Additional operations include further rounds of cell-substrate or cell-cell adhesion, as well as transferring the patterns formed by DPAC to an embedding hydrogel for long-term culture. Previously, methods for patterning oligonucleotides on surfaces and decorating cells with DNA sequences required specialized equipment and custom DNA synthesis, respectively. We report an updated version of the protocol, utilizing an inexpensive benchtop photolithography setup and commercially available cholesterol modified oligonucleotides (CMOs) deployed using a modular format. CMO-labeled cells adhere with high efficiency to DNA-patterned substrates. This approach can be used to pattern multiple cell types at once with high precision and to create arrays of microtissues embedded within an extracellular matrix. Advantages of this method include its high resolution, ability to embed cells into a three-dimensional microenvironment without disrupting the micropattern, and flexibility in patterning any cell type.

Here we present a protocol to micropattern cells at single-cell resolution using DNA-programmed adhesion. This protocol uses a benchtop photolithography platform to create patterns of DNA oligonucleotides on a glass slide and then labels cell membranes with commercially available complementary oligonucleotides. Hybridization of the oligos results in programmed cell adhesion.

DNAを用いた細胞のシンプル、手頃な価格、モジュール式パターニング

The relative positioning of cells is a key feature of the microenvironment that organizes cell-cell interactions. To study the interactions between cells ...

一本鎖DNAタイルから複雑な2次元形状の自己集合

要約

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この動画の章

折り紙インスパイアパターン化され、再構成可能な粒子の自己組織化

DNA折り紙からバイオ応答ロボットの折りたたみとキャラクタリゼーション

DNA折り紙解析と実験のためのマイカとシリコン基板の作製

DNA折り紙からバイオ応答ロボットの設計

ナノ構造構築小さな環状 DNA 分子から安定した DNA モチーフ、1 D および 2 D

センシング用途のための無機構造のDNA折り紙媒介基板ナノパターニング

情報を持つペプトイドの合成と、その配列指向動的共有自己集合

再構成可能な DNA アコーディオンラックの設計と合成

有機溶媒混合物中の複雑なナノ構造へのガンマ修飾ペプチド核酸の自己集合

DNAを用いた細胞のシンプル、手頃な価格、モジュール式パターニング

一本鎖DNAタイルから複雑な2次元形状の自己集合

要約

さらに動画を探す

この動画の章

折り紙インスパイアパターン化され、再構成可能な粒子の自己組織化

DNA折り紙からバイオ応答ロボットの折りたたみとキャラクタリゼーション

DNA折り紙解析と実験のためのマイカとシリコン基板の作製

DNA折り紙からバイオ応答ロボットの設計

ナノ構造構築小さな環状 DNA 分子から安定した DNA モチーフ、1 D および 2 D

センシング用途のための無機構造のDNA折り紙媒介基板ナノパターニング

情報を持つペプトイドの合成と、その配列指向動的共有自己集合

再構成可能な DNA アコーディオン ラックの設計と合成

有機溶媒混合物中の複雑なナノ構造へのガンマ修飾ペプチド核酸の自己集合

DNAを用いた細胞のシンプル、手頃な価格、モジュール式パターニング

再構成可能な DNA アコーディオンラックの設計と合成