A kind of origami structure based on double-stranded nucleic acid and its preparation method and application
A double-stranded nucleic acid, origami structure technology, applied in biochemical equipment and methods, genetic engineering, resistance to vector-borne diseases, etc., can solve problems such as limiting wide application, achieve good binding effect, simple preparation process, and strong versatility Effect
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Embodiment 1
[0304] In this example, an origami structure based on double-stranded nucleic acid is prepared. The schematic diagram of the preparation method is as follows: figure 1 shown, the specific steps are as follows:
[0305] (1) Expression and purification of gene editing proteins
[0306] The gene editing protein includes two parts, inactive Cas9 and inactive Cas12a. The preparation method is as follows:
[0307] The coding genes of inactive Cas9 and inactive Cas12a were constructed into the pET 28a expression vector, and the sequences were verified by sequencing; then the expression plasmids were transformed into Escherichia coli Rosetta strain, and isopropyl-β-D-thiol was added. Galactoside induces the expression of gene editing protein; collect the bacteria expressing gene editing protein and carry out ultrasonic breakage, the supernatant is preliminarily purified by affinity chromatography nickel column, and then purified by molecular sieve gel chromatography, and the purified...
Embodiment 2
[0318] In this example, the gene editing complex containing the gRNA shown in SEQ ID NO: 29-46 and the linearized P3487 (SEQ ID NO: 2) with a length of 3487 bp are used to assemble the origami structure. The method is the same as that in Example 1. Combined with schematics such as Image 6 shown;
[0319] Atomic force microscopy was performed on the origami structure, and the results were as follows Figure 7 As shown, a double-stranded nucleic acid-based origami structure was successfully assembled. The structure has a regular morphology, a particle size of about 160 nm, and good dispersibility.
Embodiment 3
[0321] Compared with Example 2, the framework DNA was P3487 without linearization treatment, and other conditions were the same as Example 2.
[0322] Atomic force microscopy was performed on the origami structure, and the results were as follows Figure 8 As shown, a double-stranded nucleic acid-based origami structure was successfully assembled. The structure has a regular morphology, a particle size of about 160 nm, and good dispersibility.
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