Ce travail a été financé par le Bureau du Programme de recherche navale Young Investigator N000141110914 Award, Office of Naval Research Grant N000141010827, NSF CAREER Award CCF1054898, New Innovator Award 1DP2OD007292 de directeur des NIH et l&#39;Institut Wyss pour Biologiquement Inspirée Fonds de démarrage Faculté de génie (PY) et Tsinghua de Pékin-Centre pour le Fonds de démarrage sciences de la vie (BW).

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The authors declare competing financial interests.

Dans l&#39;étape de formation de la structure, il est important de maintenir une concentration appropriée de cations de magnésium (par ex., 15 mM) dans le mélange de brin d&#39;ADN de nanostructures d&#39;ADN auto-assembler. De même, dans l&#39;étape de caractérisation gel d&#39;agarose / purification, il est important de maintenir une concentration de cations de magnésium approprié (par ex., 10 mM) dans le gel et le tampon du gel de fonctionnement pour retenir les nanostructures d&#39;ADN dur...

Acide nucléique précédent travail d&#39;auto-assemblage 1-25 a conduit à la construction réussie d&#39;une variété de structures complexes, y compris l&#39;ADN 2 - 5,8,10 - 13,17,23 ou d&#39;ARN 7,22 3,4,7 périodique, 22 et algorithmique 5 bidimensionnelles treillis, rubans 10,12 et tubes 4,12,13, cristaux 3D 17, 11 et polyèdres finis, formes 2D 7,8. Une méthode particulièrement efficace est échafaudée l&#39;ADN origami, par lequel un seul brin d&#39;échafaudage est plié par de nombreux courts brins de base auxiliaires pour former une forme complexe 9,14 - 16,18 - 21,25. Nous avons récemment rapporté une méthode pour construire des nanostructures discrètes avec des formes 2D prescrites en utilisant les tuiles simple brin (SST), et démontré la complexité des structures comparables à l&#39;ADN origami 26. Cette article est une adaptation de notre travail antérieur 26 et décrit des protocoles détaillés pour organiser SST adressables individuellement dans des formes 2D finis sophistiqués avec des dimensions précisément prescrites (largeurs et longueurs) et morphologies. Un avantage clé de la méthode SST est sa modularité. Chaque SST composant d&#39;une structure sert d&#39;unité de construction modulaire dans l&#39;assemblée, et les différents sous-ensembles de ces SST produire des formes distinctes. Ainsi, nous avons créé une plate-forme générale pour construire des nanostructures avec des tailles et des formes prescrites à partir de courts brins d&#39;ADN synthétiques. SST contiennent quatre domaines, chacun 10 ou 11 nucléotides de long (figure 1A). Les SST lient de telle sorte que leurs hélices parallèles créent un réseau d&#39;ADN maintenus ensemble par des liens croisés. Chaque croisement est le phosphate entre les domaines 2 et 3. Le phosphate est étiré artificiellement dans les schémas pour la clarté visuelle. Les croisements sont espacés deux spires hélicoïdales (21 bases) d&#39;intervalle (&lt;strong&gt; Figure 1B). Les rectangles composites sont désignés par leurs dimensions dans le nombre d&#39;hélices et les spires hélicoïdales. Par exemple, un rectangle qui est six hélices de large et huit spires hélicoïdales longue est référencé comme un rectangle 6H × 8T. SST peut être omis, ajoute, ou autrement réarrangés pour créer des structures de formes et de tailles (figure 1C) arbitraires. Par exemple, une forme rectangulaire peut être enroulée dans un tube à une longueur souhaitée et du rayon (figure 1D). Alternativement, le SST réseau rectangulaire peut être considérée comme une toile moléculaire constitué de SST pixels, chaque 3 nm de 7 nm. Dans cette étude, nous utilisons une toile moléculaire de 310 pleine longueur SST internes, 24 SST-métrages qui composent les limites gauche et droite, et 28 demi-longueur SST constituent les limites supérieure et inférieure. La toile a 24 doubles hélices reliées par des croisements et chaque hélice contient 28 spires hélicoïdales (294 bases) et est donc appeléune 24H × 28T toile rectangulaire. Le 24H × toile 28T a un poids moléculaire similaire à celle d&#39;une structure de l&#39;ADN origami créé à partir d&#39;un phage M13 échafaudage.

<table border="0" cellpadding="0" cellspacing="0" width="919">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Name of Material/ Equipment</td>
			<td style="width:183px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:401px;">Comments/Description</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">DNA Strands&#160;</td>
			<td style="width:183px;">Integrated DNA Technology</td>
			<td style="width:119px;"></td>
			<td>Section 3.1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">SYBR Safe DNA gel stain</td>
			<td style="width:183px;">Invitrogen</td>
			<td style="width:119px;">S33102</td>
			<td>Section 3.4.2</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Freeze&#39;N Squeeze DNA Gel Extraction Spin Columns</td>
			<td style="width:183px;">BIO-RAD</td>
			<td style="width:119px;">731-6166</td>
			<td>Section 3.6</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Bruker&#39;s Sharp Nitride Lever Probes</td>
			<td style="width:183px;">Bruker AFM Probes</td>
			<td style="width:119px;">SNL10</td>
			<td>Section 4.3</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Safe Imager 2.0 Blue Light Transilluminator</td>
			<td style="width:183px;">Invitrogen</td>
			<td style="width:119px;">G6600</td>
			<td>Section 3.6</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Centrifuge 5430R</td>
			<td style="width:183px;">Eppendorf</td>
			<td style="width:119px;">5428 000.414</td>
			<td>Section 3.6</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Transmission Electron Microscope&#160;</td>
			<td style="width:183px;">Jeol</td>
			<td style="width:119px;">Jem 1400</td>
			<td>Section 7.4</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Multimode 8</td>
			<td style="width:183px;">Veeco</td>
			<td style="width:119px;"></td>
			<td>Section 4</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Typhoon FLA 9000 Laser Scanner</td>
			<td style="width:183px;">GE Heathcare Life Sciences</td>
			<td style="width:119px;">28-9558-08</td>
			<td>Section 3.5</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Ultrapure Distilled water</td>
			<td style="width:183px;">Invitrogen</td>
			<td style="width:119px;">10977-023</td>
			<td>Section 3.7.1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Mica disk</td>
			<td style="width:183px;">SPI Supplies</td>
			<td style="width:119px;">12001-26-2</td>
			<td>Section 4.1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Steel mounting disk</td>
			<td style="width:183px;">Ted Pella, Inc.</td>
			<td style="width:119px;">16218</td>
			<td>Section 4.1</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">carbon coated copper grid for TEM</td>
			<td style="width:183px;">Electron Microscopy Sciences</td>
			<td style="width:119px;">FCF400-Cu</td>
			<td>Section 7.2</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">tweezers</td>
			<td style="width:183px;">Dumont</td>
			<td>0203-N5AC-PO</td>
			<td>Section 7.31</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">glow discharge system</td>
			<td style="width:183px;">Quorum Technologies</td>
			<td style="width:119px;">K100X</td>
			<td>Section 7.2</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">DNA Engine Tetrad 2 Peltier Thermal Cycler</td>
			<td style="width:183px;">BIO-RAD</td>
			<td>PTC&#8211;0240G</td>
			<td>Section 3.3</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Owl Easycast B2 Mini Gel Electrophoresis Systems</td>
			<td style="width:183px;">ThermoScientific</td>
			<td>B2</td>
			<td>Section 3.4.3</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Seekam LE Agarose 500G</td>
			<td style="width:183px;">Lonza</td>
			<td style="width:119px;">50004</td>
			<td>Section 3.4.1</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">GeneRuler 1kb Plus DNA Ladder, Ready-To-Use 75-20000bp</td>
			<td style="width:183px;">ThermoScientific</td>
			<td style="width:119px;">SM1333</td>
			<td>Section 3.4.4</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Nanodrop 2000c UV-vis Spectrophotometer</td>
			<td style="width:183px;">ThermoScientific</td>
			<td style="width:119px;"></td>
			<td>Section 3.7</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">0.2 um filter</td>
			<td style="width:183px;">Corning Inc.</td>
			<td style="width:119px;">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.

L&#39;auto-assemblage de SST (figure 1) donnera une 24H x 28T rectangle, comme illustré sur la Figure 2. Les séquences d&#39;ADN pour les différents TDDS peuvent être modifiés / optimisé pour permettre l&#39;étiquetage de la streptavidine (figure 3 et 4), la transformation d&#39;un rectangle dans un tube (figure 5), l&#39;auto-assemblage programmable de SST pour former des tubes et des rectangles de différentes tailles (...

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.

Auto-assemblage de formes bidimensionnelles complexes de l&#39;ADN simple brin Tuiles

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

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11.4K vues.  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.