CN111549102A - Amphiphilic DNA nano micelle and preparation method and application thereof - Google Patents
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Abstract
The invention belongs to the technical field of biological fluorescence analysis, and particularly relates to an amphiphilic DNA nano micelle and a preparation method and application thereof. The preparation method comprises the following steps: designing a specific DNA hairpin sequence H1-5 of a target miRNA, modifying a hydrophobic C12 alkane chain at the tail end of H3 to prepare sp-H3 freeze-dried powder, preparing the obtained sp-H3 freeze-dried powder into sp-H3 dispersion liquid with a hairpin structure, adding a fluorescent dye into the dispersion liquid, and performing ultrasonic treatment to obtain the DNA nano micelle. Also provides the application of the DNA micelle nano in label-free detection of target miRNA. The DNA micelle constructed by the invention not only can be rapidly assembled into a spherical nano structure, but also has larger solid-supported capacity; meanwhile, the method has the characteristics of good selectivity, convenience in operation and the like, and has important significance for measuring the biological small molecules.
Description
Technical Field
The invention belongs to the field of biological fluorescence analysis, and particularly relates to an amphiphilic DNA nano micelle and a preparation method and application thereof.
Background
The DNA nanostructure attracts attention as a highly controllable emerging material in the field of nanotechnology, and has great development potential in the aspects of drug delivery, disease diagnosis, early detection of tumor markers and the like. In recent years, various DNA nanostructures, such as DNA tetrahedrons, DNA cubes, etc., are designed as nanocages for immobilizing small molecules for application in highly sensitive analytical detection. However, the DNA sequence for assembling the nanostructure usually takes a lot of time to elaborate, and the conditions for assembling the nanostructure by the long DNA chain are harsh, and the problems of long assembling time, low assembling efficiency, etc. limit the wide application of the DNA assembly-based nanomaterial. In view of these problems, it is important to construct a DNA nanostructure with simple base sequence design and high assembly efficiency.
In recent years, it has been reported that an amphiphilic block copolymer can form a nano micelle by self-assembly, and a large amount of functional small molecules can be encapsulated in the micelle, thereby being a popular research object in the fields of constructing drug delivery systems, nano reactors and the like. However, the conventional amphiphilic copolymer micelle releases inner molecules through ultraviolet or pH adjustment, and the adjustment modes not only can not directly realize specificity and quantitative detection on biological molecules, but also can influence the stability of a reaction system through long-term ultraviolet radiation or pH environment change, thereby limiting the application value to a certain extent.
Disclosure of Invention
In view of the above, an object of the present invention is to provide an amphiphilic DNA nano-micelle. The second purpose of the invention is to provide a preparation method of the amphiphilic DNA nano-micelle, the preparation method can enable the amphiphilic DNA to be rapidly assembled into a spherical nano-structure, and the formed nano-micelle has larger solid-supported capacity. The invention also aims to provide a method for detecting target miRNA by amphiphilic DNA nano-micelle in a label-free manner, which has the characteristics of good selectivity, convenience in operation and the like. The fourth purpose of the invention is to provide an application of the amphiphilic DNA nano micelle in miRNA detection in tumor cells. In order to achieve the purpose, the invention provides the following technical scheme:
1. a preparation method of amphiphilic DNA nano-micelle comprises the following steps:
(1) designing a specific DNA hairpin sequence H1-5 of a target miRNA, wherein the nucleotide sequence is shown as SEQ ID NO: 1-5, wherein the target miRNA is miRNA-21, and the nucleotide sequence is shown as SEQ ID NO: 6, modifying a hydrophobic C12 alkane chain at the tail end of the H3 hairpin sequence to prepare amphiphilic DNA copolymer sp-H3 freeze-dried powder;
(2) preparing dispersion liquid of sp-H3 with a hairpin structure from the sp-H3 freeze-dried powder obtained in the step (1);
(3) and (3) adding a fluorescent dye into the dispersion liquid obtained in the step (2), and performing ultrasonic treatment.
As one of the preferred technical solutions: in the step (2), the dispersion is prepared by the following method: dissolving the sp-H3 lyophilized powder in a solution containing Mg2+The buffer solution is incubated for 5-10min at the temperature of 95 +/-2 ℃, and then the temperature is reduced to 25 ℃ at the speed of 0.5 ℃/min and then kept for 5-10 min.
As one of the preferred technical solutions: in the step (3), the molar ratio of the fluorescent dye to the sp-H3 with the hairpin structure in the dispersion liquid is 10: 1.
As one of the preferred technical solutions: the fluorescent dye is hydrophobic fluorescent dye, and the hydrophobic fluorescent dye is but not limited to perylene, cyanine, oil red or nile red.
As one of the preferred technical solutions: the ultrasonic power is 30-60W, and the time is 5-10 min.
2. An amphiphilic DNA nano micelle.
3. A method for detecting target miRNA (micro ribonucleic acid) by amphiphilic DNA nano-micelle in a label-free manner comprises the following steps:
adding the amphiphilic DNA nano micelle into a mixed solution of H1, H2, H4 and H5 to form a reaction solution, adding a target object miRNA to be detected into the reaction solution for incubation, and finally detecting the fluorescence response intensity.
As one of the preferred technical solutions: the incubation temperature is 35-40 ℃, and the incubation time is 90-120 min.
As one of the preferred technical solutions: the molar ratio of the amphiphilic DNA nano micelle, H1, H2, H4 and H5 in the reaction solution is 1:1:1: 1.
4. An application of amphiphilic DNA nano-micelle in miRNA detection in tumor cells.
The invention has the beneficial effects that:
(1) the constructed DNA nano micelle structure can be rapidly assembled through non-covalent action, so that the assembly time of the DNA nano structure is greatly shortened;
(2) the constructed DNA micelle is used as a novel nano-carrier, can quickly wrap a large number of related small molecules, and improves the immobilization capacity;
(3) the sp-H3 of the hairpin structure in the DNA nano micelle endows the DNA nano micelle with excellent specificity, accuracy and codeability, and provides a potential basis for realizing qualitative and quantitative analysis of a target object;
(4) the DNA nano-micelle has higher sensitivity, excellent selectivity and good reproducibility for detecting the target miRNA.
Drawings
FIG. 1 is a schematic diagram of the principle of label-free detection of target miRNA by DNA nanomicelle;
FIG. 2(A) is a fluorescence response curve of DNA nano-micelle to miRNA-21 standard solutions of different concentrations, and (B) is a standard curve of fluorescence signal response intensity value and logarithm of miRNA-21 concentration;
FIG. 3(A) is a result graph of the response sensitivity of the amphiphilic DNA nano-micelle to miRNA-21, and (B) is a result graph of the detection of miRNA-21 in cells.
Detailed Description
Preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings.
Example 1
Preparation of amphiphilic DNA nano-micelle
The miRNA of the embodiment is miRNA-21
1) Designing a specific DNA hairpin sequence H1-H5 of a target miRNA-21(SEQ ID NO: 1-5), modifying hydrophobic C12 alkane chain (Spacer C12) at the tail end of the hairpin sequence of H3 by a specific chemical coupling technology10To obtainTo amphiphilic DNA copolymers sp-H3; the freeze-dried powder of H1, H2, H4, H5 and sp-H3 is obtained by the synthesis of trivalent phosphorus chemical technology.
2) The preparation method of the amphiphilic DNA copolymer sp-H3 dispersion liquid with the hairpin structure comprises the following specific steps:
firstly, centrifuging sp-H3 lyophilized powder for 10min at 12000r/min, and then dissolving the powder in a solvent containing Mg2+The resulting mixture was uniformly dispersed in TE buffer (5 nM) to obtain a sp-H3 solution. And subpackaging the sp-H3 solution into 100 mu L EP tubes, placing the tubes in a PCR instrument for incubation at 95 ℃ for 10min, then reducing the temperature to 25 ℃ at the speed of 0.5 ℃/min, and keeping the temperature for 10min to obtain the dispersion liquid of sp-H3 with the hairpin structure.
3) Preparation of amphiphilic DNA nano-micelle: adding perylene into the obtained dispersion liquid, wherein the molar ratio of perylene to sp-H3 with hairpin structures in the dispersion liquid is 10:1, and carrying out ultrasonic treatment for 5min at the power of 50W to obtain the amphiphilic DNA nano micelle.
Example 2
Unmarked detection of amphiphilic DNA nano micelle on miRNA-21
The principle of the DNA nano-micelle label-free detection of miRNA-21(SEQ ID NO: 6) is shown in figure 1, an amphiphilic DNA copolymer sp-H3 is obtained by modifying a hydrophobic C12 alkane chain at the tail end of hydrophilic DNA, sp-H3 is formed into a hairpin structure for wrapping a fat-soluble dye perylene, and the stable DNA nano-micelle is assembled through the hydrophilic and hydrophobic interaction between the two. In the presence of a target miRNA-21, a catalytic hairpin self-assembly reaction (CHA) between DNA hairpins H1 and H2 is triggered, so that the cyclic utilization of a trace target is realized, and a large amount of H1-H intermediates are output; the intermediate can open sp-H3 on the DNA nano micelle and expose the sticky end of sp-H3. Furthermore, the sticky end of the sp-H3 can initiate a Hybridization Chain Reaction (HCR) between H4 and H5, so that the hydrophilicity of the DNA copolymer sp-H3 is enhanced, the stability of the DNA nano micelle is damaged, the fluorescent dye perylene inside the micelle is released into an organic phase, and the fluorescent signal in the aqueous phase is reduced. In a homogeneous reaction system, the intensity of the decrease of the fluorescence signal in the aqueous solution is in direct proportion to the logarithmic value of the concentration of the target miRNA-21.
1) Preparation of miRNA-21 Standard Curve
The amphiphilic DNA nano-micelle prepared in the example 1 is added into a mixed solution of H1, H2, H4 and H5 to form a reaction solution, and the molar ratio of the amphiphilic DNA nano-micelle, H1, H2, H4 and H5 in the reaction solution is 1:1:1:1: 1. Obtaining miRNA-21 freeze-dried powder by a gene synthesis technology, preparing the miRNA-21 freeze-dried powder into mother liquor with the concentration of 1 mu M, respectively adding the mother liquor into reaction solution to enable the final concentration of miRNA-21 to be 0.02nM, 0.05nM, 0.1nM, 0.2nM, 0.5nM, 1nM, 2nM, 5nM and 20nM respectively, incubating for 90min at 35 ℃, after the reaction is finished, respectively sucking 50 mu L of aqueous phase solution into a quartz cuvette by using a pipette gun, and detecting a fluorescence signal in the solution by using an F-7000 fluorescence spectrophotometer. The excitation wavelength of the fluorometer is 440nm, the collected emission spectrum has a wavelength range of 465nm to 615nm, and the slit width is 10 nm. As shown in A in FIG. 2, as the concentration of the target miRNA-21 increases from 0.02nM to 20nM, the fluorescence signal in the solution gradually decreases; as shown in B in FIG. 2, the fluorescence signal response has a good linear correlation with the logarithm of the miRNA-21 concentration, and the linear regression equation is as follows: i ═ 0.2032lg cmiRNA-21-0.4098, linear correlation coefficient r 0.9955; wherein I ═ F-F0)/F0F and F0The fluorescence response intensities of the experimental and blank groups are represented, respectively. The linear range was 20pM to 20nM with a detection limit of 6.9 pM.
2) Response sensitivity of amphiphilic DNA micelle nano to miRNA-21
Preparing a reaction solution mixed by amphiphilic DNA nano-micelles, H1, H2, H4 and H5 according to the method of the step 1), and obtaining miRNA-155(SEQ ID NO: 7) miRNA-122(SEQ ID NO: 8) miRNA-182-5p (SEQ ID NO: 9) miRNA-141(SEQ ID NO: 10) and miRNA-21 lyophilized powder, wherein miRNA-155, miRNA-122, miRNA-182-5p and miRNA-141 are used as interfering miRNAs, miRNA-21 is used as a target miRNA to be detected, the miRNA lyophilized powder is respectively added into a reaction solution to enable the concentration of each interfering miRNA to be 20nM and the concentration of the target miRNA to be 2nM, then all the miRNA lyophilized powder is simultaneously added into another reaction solution, the concentration of each interfering miRNA is 20nM and the concentration of miRNA-21 is 2nM, and incubation and fluorescence signal detection are carried out according to the conditions in the step 1). The result is shown in a in fig. 3, the fluorescence response of the solution after incubation of interfering mirnas with the concentrations of 20nM is not obvious; and the fluorescence response of the solution after the incubation of the target miRNA-21 with the concentration of 2nM is obviously enhanced. Meanwhile, after the interfering miRNA with the concentration of 20nM and the miRNA-21 with the concentration of 2nM are incubated together, the fluorescence response intensity of the solution is not obviously changed compared with the fluorescence response intensity of the solution after the incubation of the miRNA-21 with the concentration of 2 nM. These results prove that the constructed amphiphilic DNA nano-micelle has good response sensitivity to the target miRNA.
3) Method for detecting miRNA-21 content in cells by using amphiphilic DNA nano micelle
Preparing a reaction solution of amphiphilic DNA nano-micelle, H1, H2, H4 and H5 mixed according to the method of the step 1), culturing MCF-7 cells and Hela cells, collecting and counting the cells, wherein the number of the cells is 1 × 106～1×107Respectively extracting cell RNA by using cell RNA extraction kit (cargo number: RC101) to obtain RNA solutions of MCF-7 and Hela cells, diluting the two RNA solutions to 1 × 10 according to cell number350 mu L of each of the two-phase mixture, 5 × 10350 mu L of the powder/1 × 10450 μ L of each of the two drugs, 5 × 10450 μ L, and 1 × 10550 μ L of the total amount of the cells, 10 μ L of the total amount of the cells were added to 100 μ L of the reaction solution, and incubation and detection of fluorescent signal were performed under the conditions of step 1) and the results are shown in B of FIG. 3, where the number of cells was 1 × 10 according to MCF-73Increase 1 × 105The response change of the fluorescence signal is obvious; however, as the cell number of Hela gradually increases, the response of the fluorescence signal is changed slowly, and the change intensity of the overall response is low; thereby indicating that the expression level of the miRNA-21 in the MCF-7 cells is higher relative to the expression level of the miRNA-21 in the Hela cells. The result shows that the constructed amphiphilic DNA nano micelle can be used for detecting miRNA-21 in cell lysate.
Finally, the above embodiments are only intended to illustrate the technical solutions of the present invention and not to limit the present invention, and although the present invention has been described in detail with reference to the preferred embodiments, it will be understood by those skilled in the art that modifications or equivalent substitutions may be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions, and all of them should be covered by the claims of the present invention.
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Claims (10)
1. A preparation method of amphiphilic DNA nano-micelle is characterized by comprising the following steps:
(1) designing a specific DNA hairpin sequence H1-5 of the target miRNA, wherein the nucleotide sequence is shown as SEQ ID NO: 1-5, wherein the target miRNA is miRNA-21, and the nucleotide sequence is shown as SEQ ID NO: 6, modifying a hydrophobic C12 alkane chain at the tail end of the H3 hairpin sequence to prepare amphiphilic DNA copolymer sp-H3 freeze-dried powder;
(2) preparing dispersion liquid of sp-H3 with a hairpin structure from the sp-H3 freeze-dried powder obtained in the step (1);
(3) and (3) adding a fluorescent dye into the dispersion liquid obtained in the step (2), and performing ultrasonic treatment.
2. The method according to claim 1, wherein in the step (2), the dispersion is prepared as follows: dissolving the sp-H3 lyophilized powder in a solution containing Mg2+The buffer solution is incubated for 5-10min at the temperature of 95 +/-2 ℃, and then the temperature is reduced to 25 ℃ at the speed of 0.5 ℃/min and then kept for 5-10 min.
3. The method according to claim 1, wherein in the step (3), the molar ratio of the fluorescent dye to the sp-H3 having a hairpin structure in the dispersion liquid is 10: 1.
4. The method of claim 1, wherein in step (3), the fluorescent dye is a hydrophobic fluorescent dye, and the hydrophobic fluorescent dye is but not limited to perylene, cyanine, oil red or nile red.
5. The method according to claim 1, wherein the ultrasonic power in the step (3) is 30-50W for 5-10 min.
6. An amphiphilic DNA nanomicelle prepared by the method of any one of claims 1-5.
7. The method for label-free detection of target miRNA by amphiphilic DNA nano-micelle of claim 6, wherein the method comprises the following steps:
adding the amphiphilic DNA nano micelle into a mixed solution of H1, H2, H4 and H5 to form a reaction solution, adding a target object miRNA to be detected into the reaction solution for incubation, and finally detecting the fluorescence response intensity.
8. The method of claim 7, wherein the incubation temperature is 35-40 ℃ for 90-120 min.
9. The method of claim 7, wherein the amphiphilic DNA nanomicelles, H1, H2, H4 and H5 in the reaction solution are in a molar ratio of 1:1:1:1: 1.
10. The application of the amphiphilic DNA nano-micelle of claim 6 in the detection of miRNA in tumor cells.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114231599A (en) * | 2020-09-09 | 2022-03-25 | 北京师范大学 | Hybrid nanosphere based on metal-DNA coordination and preparation method and application thereof |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016149021A1 (en) * | 2015-03-13 | 2016-09-22 | Life Technologies Corporation | Methods, compositions and kits for small rna capture, detection and quantification |
| CN106498047A (en) * | 2016-10-21 | 2017-03-15 | 江南大学 | Method based on tetrahedral dual signal in situ detection intracellular microRNA of golden up-conversion nanoparticles |
| CN106822036A (en) * | 2016-12-15 | 2017-06-13 | 国家纳米科学中心 | Special target self-assembling polypeptide nano-carrier, drug-loading nanoparticles and preparation method |
| CN107574227A (en) * | 2017-10-10 | 2018-01-12 | 武汉大学 | A kind of method for nucleic acid analysis based on cascade hybridization chain reaction |
| CN108624620A (en) * | 2018-04-02 | 2018-10-09 | 湖南大学 | DNA nanometers of tetrad realizations are efficiently conveyed to nucleic acid and the super sensitivity detection method of miRNA |
| CN109486906A (en) * | 2018-11-16 | 2019-03-19 | 中山大学附属第五医院 | MicroRNA non-enzymatic amplification detection method intracellular based on the affine nanometer transport vehicle of electrostatic and cell imaging |
| CN109913534A (en) * | 2019-01-24 | 2019-06-21 | 中国药科大学 | A method of two kinds of miRNA based on concatenated dna chain circular response and fluorescence resonance energy transfer effect are detected simultaneously |
| CN110257520A (en) * | 2019-07-04 | 2019-09-20 | 福州大学 | A technique for utilizing miRNA in short capillary high speed electrophoresis detection lung carcinoma cell |
| CN110455764A (en) * | 2019-08-29 | 2019-11-15 | 青岛科技大学 | The detection method of tumour cell marker miRNA-21 and tumour cell |
-
2020
- 2020-05-28 CN CN202010470726.7A patent/CN111549102B/en active Active
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016149021A1 (en) * | 2015-03-13 | 2016-09-22 | Life Technologies Corporation | Methods, compositions and kits for small rna capture, detection and quantification |
| CN107429296A (en) * | 2015-03-13 | 2017-12-01 | 生命技术公司 | Method, composition and the kit of capture, detection and quantitative tiny RNA |
| CN106498047A (en) * | 2016-10-21 | 2017-03-15 | 江南大学 | Method based on tetrahedral dual signal in situ detection intracellular microRNA of golden up-conversion nanoparticles |
| WO2018072745A1 (en) * | 2016-10-21 | 2018-04-26 | 江南大学 | Method for double-signal in-situ detection of intracellular microrna based on gold-upconversion nanoparticle tetrahedron |
| CN106822036A (en) * | 2016-12-15 | 2017-06-13 | 国家纳米科学中心 | Special target self-assembling polypeptide nano-carrier, drug-loading nanoparticles and preparation method |
| CN107574227A (en) * | 2017-10-10 | 2018-01-12 | 武汉大学 | A kind of method for nucleic acid analysis based on cascade hybridization chain reaction |
| CN108624620A (en) * | 2018-04-02 | 2018-10-09 | 湖南大学 | DNA nanometers of tetrad realizations are efficiently conveyed to nucleic acid and the super sensitivity detection method of miRNA |
| CN109486906A (en) * | 2018-11-16 | 2019-03-19 | 中山大学附属第五医院 | MicroRNA non-enzymatic amplification detection method intracellular based on the affine nanometer transport vehicle of electrostatic and cell imaging |
| CN109913534A (en) * | 2019-01-24 | 2019-06-21 | 中国药科大学 | A method of two kinds of miRNA based on concatenated dna chain circular response and fluorescence resonance energy transfer effect are detected simultaneously |
| CN110257520A (en) * | 2019-07-04 | 2019-09-20 | 福州大学 | A technique for utilizing miRNA in short capillary high speed electrophoresis detection lung carcinoma cell |
| CN110455764A (en) * | 2019-08-29 | 2019-11-15 | 青岛科技大学 | The detection method of tumour cell marker miRNA-21 and tumour cell |
Non-Patent Citations (3)
| Title |
|---|
| MEIJUAN LIANG等: "Electrochemical Biosensor for MicroRNA Detection Based on Cascade Hybridization Chain Reaction", 《CHEMELECTROCHEM》 * |
| XIN PENG等: "Rapid self-disassembly of DNA diblock copolymer micelles via target induced hydrophilic-hydrophobic regulation for sensitive MiRNA detection", 《CHEMCOMM》 * |
| 汤玉娇等: "基于DNA模板点击化学和催化发夹型DNA自组装反应的新型均相电化学生物传感器检测miRNA-21", 《分析化学》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114231599A (en) * | 2020-09-09 | 2022-03-25 | 北京师范大学 | Hybrid nanosphere based on metal-DNA coordination and preparation method and application thereof |
| CN114231599B (en) * | 2020-09-09 | 2023-07-18 | 北京师范大学 | Hybrid nanosphere based on metal-DNA coordination and preparation method and application thereof |
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