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Step-by-step assembly of DNA origami units

A unit group, self-assembly technology, applied in the biological field, can solve problems such as mismatches and inability to obtain designed structures

Active Publication Date: 2021-08-31
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the one-step method means that the specificity of the junction of each structural monomer is required, otherwise there will be mismatches, etc., and the designed structure cannot be obtained.

Method used

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  • Step-by-step assembly of DNA origami units
  • Step-by-step assembly of DNA origami units
  • Step-by-step assembly of DNA origami units

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0182] Embodiment 1, DNA origami unit step-by-step assembly method

[0183] 1. The core idea of ​​this method

[0184] Fix the DNA nanostructure on some kind of solid phase plane, and add DNA origami units step by step. exist figure 1 , shows a schematic diagram of the step-by-step assembly method. Each origami unit has two sticky ends, and the two sticky ends are complementary to the corresponding sticky ends of the origami units on both sides. Each origami unit follows its DNA nanostructure The sequence is numbered sequentially and added step by step.

[0185] Using magnetic beads with streptavidin (commodity is Dynabeads M-270) as the solid phase plane, the linking single strand with biotin at the 3' end is connected to the On the magnetic beads, the base of 13bp (TATGATTGAGAGATT) connected to the 5' end of the single strand is complementary to a cohesive end of the first DNA origami unit to realize the immobilization of the first DNA origami unit; then through the secon...

Embodiment 2

[0217] Embodiment 2, the limit of the number of steps to explore the structural design and 12-step structural results

[0218] 1. Structural design of origami unit step-by-step assembly method to explore the limit of steps

[0219] 1. Multiple supplementary short chains and multiple short chains with sticky ends required for the limit of the number of preparation steps to explore the structure

[0220] Such as figure 2 As shown, 28 rectangular target DNA origami units are set according to the limit of steps to explore the structure, and caDNAno software is used to design multiple supplementary short chains and multiple sticky ends according to the long-chain DNA molecular sequence of each target DNA origami unit Short chains; a plurality of short chains with sticky ends is to extend a plurality of supplementary short chain bases on the edge of each target DNA origami unit out of sticky ends to obtain multiple short chains with sticky ends;

[0221] To complete the experimen...

Embodiment 3

[0305] Example 3. Assembly of Isosceles Triangular DNA Nanostructures Using Origami Unit Step-by-Step Assembly Method

[0306] 1. Assembly of isosceles triangular DNA nanostructures by step-by-step assembly of origami units

[0307] 1. Design of the origami unit constituting the isosceles triangle DNA nanostructure and the long and short chains constituting the origami unit

[0308] In order to explore the limit of the number of monomers added to the step-by-step assembly method, an isosceles triangular DNA nanostructure is designed as Figure 4 As shown, the multiple supplementary short chains and multiple short chains with sticky ends required to prepare this structure are the same as in Example 2-1.

[0309] Such as Figure 4 As shown, first set up 21 rectangular target DNA origami units (numbered 1-21) connected sequentially according to the pre-formed isosceles triangle DNA nanostructure, and then use caDNAno software to design and synthesize each target DNA origami uni...

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Abstract

A method for assembling nucleic acid nanostructures from nucleic acid origami units step by step, comprising the following steps: 1) preparing target nucleic acid origami unit groups for the first step in the following steps 2) and 3) according to the pre-formed nucleic acid nanostructures to The m-th step target nucleic acid origami unit group, m is an integer greater than or equal to 2; 2) The target nucleic acid origami unit group obtained in step 1) is connected step by step in order and fixed on the stationary phase connected with the connecting chain to obtain the connection A nucleic acid nanostructure with connecting strands; 3) adding a separation strand that is completely complementary to the connecting strands to the nucleic acid nanostructures connected with connecting strands obtained in step 2), and performing a separation reaction to obtain a nucleic acid nanostructure. This method achieves the successful assembly of more structural monomers, showing the high self-assembly efficiency of this new method.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a nucleic acid structure and a synthesis method thereof in the field of DNA nanotechnology, in particular to a step-by-step assembly method of DNA origami units. Background technique [0002] In the 1980s, Seeman first proposed that DNA could be assembled into a complex spatial structure using the principle of DNA base complementary pairing, creating a new field of using DNA as a nanoscale construction material rather than a carrier of genetic information, and named it as DNA nanotechnology. Subsequently, by constructing different primitive modules, such as DX (double-crossover), TX (triple-crossover) modules, cross modules and symmetrical modules, and using modules to assemble various graphic structures (two-dimensional arrays, square Grids, etc.), but the modular assembly is based on the complementary base pairing of small structural units to form a larger graph structure, and its s...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12Q1/68
Inventor 魏迪明杨林枫李逸凡王雅琪
Owner TSINGHUA UNIV