Method for regulating and controlling stability of RNA (Ribonucleic Acid) molecules by negentropy water
By dissolving RNA molecules in negative entropy water for apparent modification and lyophilization, the problem of insufficient stability of RNA molecules is solved, and the high stability and wide application of RNA molecules are achieved.
Patent Information
- Application Number
- CN202411485847.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-23
- Publication Date
- 2025-03-04
AI Technical Summary
RNA molecules have low stability and are easily destroyed by environmental factors, which limits their application in the field of biomedical science.
The RNA molecules are dissolved in negative entropy water and are apparently modified, followed by lyophilization to improve the stability of the RNA molecules.
Through this method, the stability of RNA molecules is significantly improved, making their application in the field of biomedical medicine more widely and reliable.
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Figure CN119552854A_ABST
Abstract
Description
Technical Field
[0001] The present application relates to the field of biotechnology, and in particular to a method for regulating the stability of RNA molecules using negative entropy water. Background Art
[0002] RNA is an important biological molecule that participates in the transcription and translation of genetic information. However, RNA molecules have low stability and are easily destroyed by environmental factors, which limits their application in the biomedical field. Therefore, in the prior art, there is a lack of an effective method to improve the stability of RNA molecules. Summary of the invention
[0003] Based on this, it is necessary to provide a method for regulating the stability of RNA molecules with negative entropy water to solve the technical problem that the stability of RNA molecules is not high enough.
[0004] In order to solve the above technical problems, this application provides the following technical solutions: A method for regulating RNA molecule stability using negative entropy water, characterized in that the method comprises: Dissolve the RNA molecule in negative entropy water to obtain an RNA molecule solution; Performing epitope modification on the RNA molecules in the RNA molecule solution to obtain a target RNA molecule solution; The target RNA molecule solution is freeze-dried to obtain freeze-dried RNA molecules.
[0005] Furthermore, before dissolving the RNA molecules in negative entropy water to obtain the RNA molecule solution, the process further comprises: RNA is extracted and purified from tissues or cells to obtain RNA molecules.
[0006] Furthermore, the RNA extraction and purification of tissues or cells to obtain RNA molecules includes: Phenol-chloroform extraction is used to extract RNA from tissues or cells and purify them to obtain RNA molecules; or, RNA is extracted and purified from tissues or cells using column chromatography to obtain RNA molecules.
[0007] Furthermore, after freeze-drying the target RNA molecule solution to obtain freeze-dried RNA molecules, the method further comprises: The freeze-dried RNA molecules were tested to obtain various types of indicator data; According to the multiple types of indicator data, the analysis results of the freeze-dried RNA molecules are obtained.
[0008] Furthermore, the multiple types of index data include molecular weight, purity and stability indexes.
[0009] Furthermore, after obtaining the analysis results of the freeze-dried RNA molecules according to the multiple types of indicator data, the method further includes: If one or several indicators of the multiple types of indicator data do not meet the preset requirements, the method of extracting and purifying RNA from tissues or cells is changed, and / or the method of epigenetically modifying the RNA molecules in the RNA molecule solution and / or the freeze-drying conditions are adjusted.
[0010] Furthermore, the step of performing epitope modification on the RNA molecules in the RNA molecule solution to obtain the target RNA molecule solution comprises: The RNA molecules in the RNA molecule solution are modified by chemical methods to obtain a target RNA molecule solution.
[0011] Furthermore, the chemical method is used to modify the RNA molecules in the RNA molecule solution to obtain the target RNA molecule solution, including: The target RNA molecule solution is obtained by adding chemical groups to certain nucleotides of the RNA molecules in the RNA molecule solution to modify the RNA molecules in the RNA molecule solution.
[0012] Furthermore, the chemical group includes methyl and / or acetyl.
[0013] Also provided is an RNA molecule, which is obtained by regulation by any of the aforementioned regulation methods.
[0014] In the scheme provided in this application, the RNA molecules to be studied are dissolved in negative entropy water, and then the RNA molecules are modified to improve their stability. Finally, the modified RNA molecule solution is freeze-dried to obtain stable RNA molecules, which improves the stability of RNA molecules. The whole process does not produce harmful substances and is environmentally friendly. The RNA molecules prepared by this method can be widely used in biomedical fields such as drug delivery, gene therapy, diagnostic reagents, and control of the occurrence and development of cancer. At the same time, the implementation of this method does not require complex equipment and conditions, and has good practicality and broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS
[0015] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for use in the description of the embodiments of the present application will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative labor.
[0016] Figure 1 This is a schematic diagram of a process for regulating RNA molecule stability using negative entropy water in one embodiment of the present application; Figure 2 This is another schematic flow chart of a method for regulating RNA molecule stability with negative entropy water in one embodiment of the present application. DETAILED DESCRIPTION
[0017] The following will be combined with the drawings in the embodiments of the present application to clearly and completely describe the technical solutions in the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of this application.
[0018] It should be understood that RNA is an important class of biological molecules that participate in the transcription and translation of genetic information. RNA molecules can be widely used in biomedical fields such as drug delivery, gene therapy, diagnostic reagents, and the occurrence and development of cancer control, and therefore have a wide range of applications. However, the stability of RNA molecules is relatively low and is easily destroyed by environmental factors, which limits their application in the above-mentioned fields. Therefore, in the prior art, there is a lack of an effective method to improve the stability of RNA molecules. In this regard, the present application embodiment provides a method for regulating the stability of RNA molecules by negative entropy water, and the RNA molecules obtained by the regulation method, to improve the stability of RNA molecules, described below.
[0019] like Figure 1 As shown, in one embodiment, a method for regulating RNA molecule stability with negative entropy water is provided, the method comprising: S10: Extracting and purifying RNA from tissues or cells to obtain RNA molecules.
[0020] S20: dissolving the RNA molecule in negative entropy water to obtain an RNA molecule solution; S30: performing epigenetic modification on the RNA molecules in the RNA molecule solution to obtain a target RNA molecule solution; S40: freeze-drying the target RNA molecule solution to obtain freeze-dried RNA molecules.
[0021] Regarding negative entropy water, it should be understood that the negative entropy water is water obtained through negative entropy treatment technology. By treating the zero-point particle water negative entropy, the negative entropy and fine ordering of the water molecule energy are completed, forming negative entropy water with a high-energy ordered diamond mesh structure. The difference from ordinary water is that ordinary water is positive entropy water or negative water, and its water molecule structure is composed of two positively charged hydrogen atoms and one negatively charged oxygen atom. Ordinary water is measured by an oscilloscope and presents a disordered linear structure waveform; however, the negative entropy water used in the embodiment of the present application is positive water, and its water molecule structure is composed of two negatively charged hydrogen atoms and one positively charged oxygen atom. The negative entropy water is measured by an oscilloscope and presents an ordered diamond mesh structure waveform.
[0022] In addition, in addition to the functions of ordinary water, the unique feature of negative entropy water is that it can quickly eliminate the entropy increase state of the life system and restore it to a negative entropy healthy state. In other words, in the process of life, as the effective energy in the water gradually dissipates and degenerates, it can bring more disorder and invalid energy to the entire life system, causing entropy stagnation in the life body. Negative entropy water is to holographically transform water into negative entropy, empower it without loss in the process of life operation, and quickly transform the chaos and invalid energy of the life system into orderly effective energy, reversing the negative entropy phenomenon of the body's life in time and space. Ordinary water brings irreversible entropy increase state to the life system while nourishing life, while negative entropy water empowers the life system while nourishing life, bringing a reversible negative entropy state. This is the biggest difference between negative entropy water and all known water on the earth. It can be seen that the differences between negative entropy water and ordinary water include different energy, different structure, different waveform and different function. Using negative entropy water is a true life partner that integrates safety, health and function, which meets the current needs of green development.
[0023] Therefore, in this embodiment, the RNA molecule to be studied is dissolved in negative entropy water, and the special physical properties of negative entropy water are used to improve the stability of the RNA molecule through a specific preparation method. That is, after dissolving in negative entropy water, the RNA molecule is then subjected to surface modification to improve its stability. Finally, the modified RNA molecule solution is freeze-dried to obtain a stable RNA molecule, which improves the stability of the RNA molecule. The whole process does not produce harmful substances and is environmentally friendly. The RNA molecules prepared by this method can be widely used in biomedical fields such as drug delivery, gene therapy, diagnostic reagents, and control of the occurrence and development of cancer. At the same time, the implementation of this method does not require complex equipment and conditions, and has good practicality and broad application prospects.
[0024] In one embodiment, step S10, that is, extracting and purifying RNA from tissues or cells to obtain RNA molecules, includes: Phenol-chloroform extraction is used to extract RNA from tissues or cells and purify them to obtain RNA molecules; or, RNA is extracted and purified from tissues or cells using column chromatography to obtain RNA molecules.
[0025] Among them, in the phenol-chloroform extraction method, the use of a mixture of phenol and chloroform can effectively separate RNA, DNA and protein in tissues or systems. In the final step of the extraction process, ethanol is added to precipitate RNA, thereby separating and purifying RNA molecules.
[0026] It should be understood that there are many ways to extract and purify RNA from tissues or cells. In this embodiment, phenol-chloroform extraction or column chromatography can be used to extract and purify RNA from tissues or cells to obtain the desired RNA molecules. The specific purification method is not limited in the embodiments of this application.
[0027] In one embodiment, if Figure 2 As shown, after freeze-drying the target RNA molecule solution to obtain freeze-dried RNA molecules, the method further comprises: S50: Detect the freeze-dried RNA molecules to obtain various types of indicator data; S60: Obtaining analysis results of the freeze-dried RNA molecules according to the multiple types of indicator data.
[0028] Exemplarily, the multiple types of index data include indicators such as molecular weight, purity and stability of RNA molecules. The above index data can be detected and analyzed by gel electrophoresis or ultraviolet spectroscopy to obtain index data such as molecular weight, purity and stability, without specific limitation.
[0029] In one embodiment, the step of performing epitope modification on the RNA molecules in the RNA molecule solution to obtain the target RNA molecule solution comprises: The RNA molecules in the RNA molecule solution are modified by chemical methods to obtain a target RNA molecule solution.
[0030] Furthermore, the method of using a chemical method to perform surface modification on the RNA molecules in the RNA molecule solution to obtain a target RNA molecule solution includes: adding chemical groups to certain nucleotides of the RNA molecules in the RNA molecule solution to perform surface modification on the RNA molecules in the RNA molecule solution to obtain a target RNA molecule solution.
[0031] Wherein, the chemical groups include methyl and / or acetyl groups.
[0032] In this embodiment, epigenetic modification is used to modify RNA molecules in solution to improve their stability, interaction with proteins and their location in cells. Chemical modification is to add specific chemical groups such as methyl and acetyl groups to certain nucleotides of RNA molecules to change the chemical properties and stability of RNA molecules. For example, methylation usually occurs on cytosine and adenine residues of RNA, which can affect the folding of RNA molecules and their interaction with proteins.
[0033] It should be noted that the above embodiments provide a chemical method for performing surface modification on the RNA molecules in the RNA molecule solution to obtain the target RNA molecule solution. In other embodiments, other surface modification methods may also be used, which are not specifically limited.
[0034] In one embodiment, after obtaining the analysis results of the freeze-dried RNA molecules based on the multiple types of indicator data, it also includes: if one or several indicators of the multiple types of indicator data do not meet the preset regulations, then changing the method of extracting and purifying RNA from tissues or cells, and / or adjusting the method of epigenetically modifying the RNA molecules in the RNA molecule solution, and / or the freeze-drying conditions.
[0035] In this embodiment, after detection and analysis, if some indicators of the freeze-dried RNA molecule do not meet the requirements, the steps in the regulation process can be changed to achieve the desired purpose. For example, if the indicators do not meet the requirements, the following regulation process of the RNA molecule can be adjusted through the following regulation actions: 1) Review the test results: First, confirm whether the indicator detection method is correctly executed and whether all instruments and reagents are operating normally.
[0036] 2) Adjust the purification process: If the purity is insufficient, you may need to reconsider the steps of RNA molecule extraction and purification, for example, replace column chromatography or phenol-chloroform extraction, or change the purification method.
[0037] 3) Optimize modification methods: If the stability is not up to standard, it may be necessary to adjust the epigenetic modification method of the RNA molecule, for example, by adding a cap to the 3' end to improve the stability of the RNA molecule.
[0038] 4) Adjust the freeze-drying conditions: If the molecular weight indicators are inconsistent, you may need to readjust the freeze-drying conditions, such as the freeze-drying temperature, time and pressure, to ensure that the RNA solution remains stable during the freeze-drying process.
[0039] 5) Repeat the experiment: If the standard is still not met after the above adjustments, it may be necessary to repeat the entire experimental process, including RNA extraction, modification and freeze-drying, until the required requirements are met.
[0040] It should be noted that during or after the adjustment process, samples that do not meet the standards can be further analyzed to explore the root cause of the problem, which may involve the quality of the original sample, the accuracy of the experimental operation, or the quality of the materials and reagents used.
[0041] In summary of the above embodiments, the present application provides a method for regulating the stability of RNA molecules by negative entropy water, dissolving the RNA molecules to be studied in negative entropy water, and utilizing the special physical properties of negative entropy water in combination with specific preparation methods such as apparent modification to improve the stability of RNA molecules, that is, after dissolving in negative entropy water, the RNA molecules are then subjected to apparent modification to improve their stability, and finally the modified RNA molecule solution is freeze-dried to obtain stable RNA molecules, thereby improving the stability of RNA molecules. The whole process does not produce harmful substances and is environmentally friendly. The RNA molecules prepared by this method can be widely used in biomedical fields such as drug delivery, gene therapy, diagnostic reagents, and control of the occurrence and development of cancer. At the same time, the implementation of this method does not require complex equipment and conditions, and has good practicality and broad application prospects.
[0042] It should be noted that the above-mentioned RNA molecules include various types, including but not limited to small interfering RNA (siRNA), etc., and are not specifically limited.
[0043] Taking the preparation of small interfering RNA (siRNA) as an example, the specific implementation method of the present application is explained. First, ordinary tap water is subjected to physical treatment processes such as filtration, degassing, and negative entropy treatment to prepare negative entropy water. Then, the small interfering RNA (siRNA) molecules to be studied are dissolved in the negative entropy water. Next, the small interfering RNA (siRNA) molecules dissolved in the negative entropy water are chemically modified to improve their stability. The modified small interfering RNA (siRNA) molecule solution is freeze-dried to obtain a stable small interfering RNA (siRNA) molecule. Finally, the freeze-dried small interfering RNA (siRNA) molecules are tested and analyzed, including indicators such as molecular weight, purity, and stability, to verify their stability.
[0044] In one embodiment, an RNA molecule is also provided, wherein the RNA molecule is obtained by any of the above-mentioned regulation methods.
[0045] The RNA molecules obtained by the control method provided in this embodiment have high stability, and the RNA molecules prepared by the method have high biocompatibility and low immunogenicity, and can be widely used in biomedical fields such as drug delivery, gene therapy, diagnostic reagents, and control of the occurrence and development of cancer. At the same time, the implementation of the method does not require complex equipment and conditions, and has good practicality and broad application prospects.
[0046] The embodiments described above are only used to illustrate the technical solutions of the present application, rather than to limit them. Although the present application has been described in detail with reference to the aforementioned embodiments, a person skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. Such modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present application, and should all be included in the protection scope of the present application.
Claims
1. A method for regulating RNA molecule stability using negative entropy water, characterized in that: The method comprises: Dissolve the RNA molecule in negative entropy water to obtain an RNA molecule solution; Performing epitope modification on the RNA molecules in the RNA molecule solution to obtain a target RNA molecule solution; The target RNA molecule solution is freeze-dried to obtain freeze-dried RNA molecules.
2. The method for regulating RNA molecule stability by negative entropy water according to claim 1, characterized in that: Before dissolving the RNA molecules in the negative entropy water to obtain the RNA molecule solution, the method further comprises: RNA is extracted and purified from tissues or cells to obtain RNA molecules.
3. The method for regulating RNA molecule stability by negative entropy water as claimed in claim 2, characterized in that: The method of extracting and purifying RNA from tissues or cells to obtain RNA molecules comprises: Phenol-chloroform extraction is used to extract RNA from tissues or cells and purify them to obtain RNA molecules; or, RNA is extracted and purified from tissues or cells using column chromatography to obtain RNA molecules.
4. The method for regulating RNA molecule stability by negative entropy water as claimed in claim 2, characterized in that: After freeze-drying the target RNA molecule solution to obtain freeze-dried RNA molecules, the method further comprises: The freeze-dried RNA molecules were tested to obtain various types of indicator data; According to the multiple types of indicator data, the analysis results of the freeze-dried RNA molecules are obtained.
5. The method for regulating RNA molecule stability by negative entropy water as claimed in claim 4, characterized in that: The various types of index data include molecular weight, purity and stability indexes.
6. The method for regulating RNA molecule stability by negative entropy water as claimed in claim 4, characterized in that: After obtaining the analysis results of the freeze-dried RNA molecules according to the multiple types of indicator data, the method further includes: If one or several indicators of the multiple types of indicator data do not meet the preset requirements, the method of extracting and purifying RNA from tissues or cells is changed, and / or the method of epigenetically modifying the RNA molecules in the RNA molecule solution and / or the freeze-drying conditions are adjusted.
7. The method for regulating RNA molecule stability using negative entropy water according to any one of claims 1 to 6, characterized in that: The method of performing epigenetic modification on the RNA molecules in the RNA molecule solution to obtain the target RNA molecule solution comprises: The RNA molecules in the RNA molecule solution are modified by chemical methods to obtain a target RNA molecule solution.
8. The method for regulating RNA molecule stability by negative entropy water as claimed in claim 7, characterized in that: The method of using a chemical method to modify the RNA molecules in the RNA molecule solution to obtain a target RNA molecule solution includes: The target RNA molecule solution is obtained by adding chemical groups to certain nucleotides of the RNA molecules in the RNA molecule solution to modify the RNA molecules in the RNA molecule solution.
9. The method for regulating RNA molecule stability by negative entropy water as claimed in claim 8, characterized in that: The chemical groups include methyl and / or acetyl groups.