Method for researching non-coding RNA (Ribonucleic Acid) regulated RNA polymerase I transcription mechanism and application of non-coding RNA regulated RNA polymerase I transcription mechanism

Abstract

The invention provides a method for researching a non-coding RNA regulation RNA polymerase I transcription mechanism and application thereof, and the method comprises the steps of non-coding RNA data collection, RNA data cleaning design and synthesis, transcription regulation mechanism, non-coding RNA annotation analysis, regulation RNA function verification, RNA expression profile analysis, polymerase I association analysis and function verification. The polymerase I transcriptional activity can be accurately regulated and controlled, a new strategy is provided for disease treatment, accurate control over cell proliferation and differentiation can be achieved through specific regulation and control over non-coding RNA, individualized treatment of cancers and hereditary diseases is made possible, some long-chain non-coding RNA molecules can participate in the transcriptional regulation and control process, and therefore the transcription activity of the polymerase I can be accurately regulated and controlled. The gene expression is further influenced by regulating and controlling the stability and splicing process of the non-coding RNA.

Description

Technical Field

[0001] The present invention belongs to the field of non-coding RNA, and particularly relates to a method for studying the mechanism of non-coding RNA regulating RNA polymerase I transcription and an application thereof. Background Art

[0002] Non-coding RNA plays an important regulatory role in the life activities of cells. It has been found that it has an important regulatory effect on the transcription of the nucleolus structure, which can deepen the understanding of the structure and function of the nucleolus, and provide a theoretical basis for the development of new treatment methods. It is of great significance for understanding the molecular mechanisms of cell growth, development and disease occurrence and development, and can inhibit the growth rate of tumors, which provides a new target for the design of anti-cancer drugs and demonstrates the potential application value of non-coding RNA in cancer treatment. RNA polymerase I is the enzyme responsible for the transcription of most ribosomal RNA in cells, and its activity is essential for cell proliferation and survival. However, non-coding RNA cannot specifically regulate polymerase I transcription, and cannot accurately regulate polymerase I transcription activity, which limits its application in disease treatment and biotechnology.

[0003] Therefore, in view of the shortcomings of the above scheme in actual production and implementation, it was revised and improved. At the same time, in the spirit and concept of seeking excellence, with the assistance of professional knowledge and experience, and after many ingenuity and experiments, the present invention was created to provide a method for studying the mechanism of non-coding RNA regulating RNA polymerase I transcription and its application, which is used to solve the problem. Summary of the invention

[0004] The present invention provides a method for studying the mechanism of non-coding RNA regulating RNA polymerase I transcription and its application, which solves the problem that non-coding RNA in the prior art cannot specifically regulate polymerase I transcription and cannot accurately regulate polymerase I transcription activity, thus limiting its application in disease treatment and biotechnology.

[0005] The technical solution of the present invention is achieved as follows: a method for studying the mechanism of non-coding RNA regulating RNA polymerase I transcription and its application, characterized in that the method steps of the RNA polymerase I transcription mechanism are as follows: (1) Collecting non-coding RNA data: In non-coding RNA research, it is necessary to collect databases and public data sets related to non-coding RNA, including the sequence, structure, function of non-coding RNA and transcriptomics data that integrates RNA expression levels. Some statistical methods and algorithms are used to reduce the dimension and standardize the data for subsequent analysis. (2) RNA data cleaning, design and synthesis: Based on the polymerase I transcription mechanism and the known RNA functions, RNA sequences with regulatory potential are designed, and the designed RNA molecules are obtained through chemical synthesis or biosynthesis. In order to better study the functions and regulatory mechanisms of non-coding RNA, data from different sources need to be integrated and analyzed; (3) Transcriptional regulatory mechanism: Using the molecular mechanism of the interaction between non-coding RNA data and polymerase I and its auxiliary factors, determine the key sites and signaling pathways of RNA data regulating polymerase I transcription, using in vitro transcription systems or cell models; (4) Annotation analysis of non-coding RNA: Annotation analysis is the process of matching unlabeled sequences with known information. In the study of non-coding RNA, it is necessary to annotate the sequence of non-coding RNA, including finding the start and end sites of non-coding RNA, and then predicting its secondary structure and functional region; (5) Verification of regulatory RNA function: Verify the regulatory effect of non-coding RNA on polymerase I transcriptional activity in cell models and animal models, evaluate its impact on cell proliferation, differentiation and disease status, and evaluate the inhibitory effect of polymerase I on cancer cell proliferation in cell models; (6) RNA expression profiling: A large amount of non-coding RNA expression data can be obtained through high-throughput sequencing. RNA data provides a series of expression profiling methods and tools to help researchers explore the potential functions and regulatory mechanisms of non-coding RNA; (7) Polymerase I association analysis: RNA analysis can be used to analyze the association between non-coding RNA and diseases. By integrating the disease database and the non-coding RNA database, the probability of association between non-coding RNA and diseases can be predicted, and correlation analysis and network analysis can be performed; (8) Functional verification: In order to further verify the function and mechanism of non-coding RNA, laboratory verification is required. Researchers design appropriate experimental plans and analyze experimental results to further verify the function and mechanism of non-coding RNA; As a preferred embodiment, the step (2) is performed by designing a non-coding RNA targeting a specific transcription factor, and by binding to the transcription factor, affecting its interaction with polymerase I, thereby achieving the purpose of mutual synthesis of different non-coding RNAs.

[0006] As a preferred embodiment, in step (3), the polymerase I transcription activity can be collected by adding the designed regulatory RNA into the in vitro transcription system and detecting the change in the RNA transcription level, thereby achieving precise regulation of the polymerase I transcription activity.

[0007] As a preferred embodiment, the step (4) predicts the secondary structure and functional region of the non-coding RNA through annotation of the non-coding RNA. The non-coding RNA data provides annotation tools and databases to help researchers annotate non-coding RNA sequences.

[0008] As a preferred embodiment, the step (7) utilizes the important regulatory role of non-coding RNA in the occurrence and development of diseases, and utilizes biological big data technology to analyze the relationship between non-coding RNA and diseases.

[0009] After adopting the above technical scheme, the beneficial effects of the present invention are as follows: the present invention can accurately regulate the transcriptional activity of polymerase I, provide a new strategy for disease treatment, and through the specific regulation of non-coding RNA, it can achieve precise control of cell proliferation and differentiation, providing the possibility for personalized treatment of cancer and genetic diseases. Some long-chain non-coding RNA molecules can participate in the transcriptional regulation process, and further affect gene expression by regulating the stability and splicing process of non-coding RNA. In addition, it can also be applied to the field of biotechnology, and by designing specific non-coding RNA molecules, it can achieve precise regulation of polymerase I transcriptional activity. This method has broad application prospects in disease treatment and biotechnology, and can provide new strategies for personalized treatment of cancer, genetic diseases and metabolic diseases. DETAILED DESCRIPTION

[0010] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. Example

[0011] A method for studying the mechanism of non-coding RNA regulating RNA polymerase I transcription and its application, characterized in that the method steps of the RNA polymerase I transcription mechanism are as follows: (1) Collecting non-coding RNA data: In non-coding RNA research, it is necessary to collect databases and public data sets related to non-coding RNA, including the sequence, structure, function of non-coding RNA and transcriptomics data that integrates RNA expression levels. Some statistical methods and algorithms are used to reduce the dimension and standardize the data for subsequent analysis. (2) RNA data cleaning, design and synthesis: Based on the polymerase I transcription mechanism and the known RNA functions, RNA sequences with regulatory potential are designed, and the designed RNA molecules are obtained through chemical synthesis or biosynthesis. In order to better study the functions and regulatory mechanisms of non-coding RNA, data from different sources need to be integrated and analyzed; (3) Transcriptional regulatory mechanism: Using the molecular mechanism of the interaction between non-coding RNA data and polymerase I and its auxiliary factors, determine the key sites and signaling pathways of RNA data regulating polymerase I transcription, using in vitro transcription systems or cell models; (4) Annotation analysis of non-coding RNA: Annotation analysis is the process of matching unlabeled sequences with known information. In the study of non-coding RNA, it is necessary to annotate the sequence of non-coding RNA, including finding the start and end sites of non-coding RNA, and then predicting its secondary structure and functional region; (5) Verification of regulatory RNA function: Verify the regulatory effect of non-coding RNA on polymerase I transcriptional activity in cell models and animal models, evaluate its impact on cell proliferation, differentiation and disease status, and evaluate the inhibitory effect of polymerase I on cancer cell proliferation in cell models; (6) RNA expression profiling: A large amount of non-coding RNA expression data can be obtained through high-throughput sequencing. RNA data provides a series of expression profiling methods and tools to help researchers explore the potential functions and regulatory mechanisms of non-coding RNA; (7) Polymerase I association analysis: RNA analysis can be used to analyze the association between non-coding RNA and diseases. By integrating the disease database and the non-coding RNA database, the probability of association between non-coding RNA and diseases can be predicted, and correlation analysis and network analysis can be performed; (8) Functional verification: In order to further verify the function and mechanism of non-coding RNA, laboratory verification is required. Researchers design appropriate experimental plans and analyze experimental results to further verify the function and mechanism of non-coding RNA; Furthermore, in step (2), a non-coding RNA targeting a specific transcription factor is designed to bind to the transcription factor, thereby affecting its interaction with polymerase I, thereby achieving the purpose of mutual synthesis of different non-coding RNAs.

[0012] Furthermore, in step (3), the polymerase I transcription activity can be collected by adding the designed regulatory RNA to the in vitro transcription system and detecting the change in the RNA transcription level, thereby achieving precise regulation of the polymerase I transcription activity.

[0013] Furthermore, the step (4) predicts the secondary structure and functional region of the non-coding RNA through annotation of the non-coding RNA. The non-coding RNA data provides annotation tools and databases to help researchers annotate non-coding RNA sequences.

[0014] Furthermore, the step (7) utilizes the fact that non-coding RNA plays an important regulatory role in the occurrence and development of diseases, and the association between non-coding RNA and diseases can be analyzed using biological big data technology. Example

[0015] A method for studying the mechanism of non-coding RNA regulating RNA polymerase I transcription and its application, characterized in that the method steps of the RNA polymerase I transcription mechanism are as follows: (1) Collecting non-coding RNA data: In non-coding RNA research, it is necessary to collect databases and public data sets related to non-coding RNA, including the sequence, structure, function of non-coding RNA and transcriptomics data that integrates RNA expression levels. Some statistical methods and algorithms are used to reduce the dimension and standardize the data for subsequent analysis. (2) RNA data cleaning, design and synthesis: Based on the polymerase I transcription mechanism and the known RNA functions, RNA sequences with regulatory potential are designed, and the designed RNA molecules are obtained through chemical synthesis or biosynthesis. In order to better study the functions and regulatory mechanisms of non-coding RNA, data from different sources need to be integrated and analyzed; (3) Transcriptional regulatory mechanism: Using the molecular mechanism of the interaction between non-coding RNA data and polymerase I and its auxiliary factors, determine the key sites and signaling pathways of RNA data regulating polymerase I transcription, using in vitro transcription systems or cell models; (4) Annotation analysis of non-coding RNA: Annotation analysis is the process of matching unlabeled sequences with known information. In the study of non-coding RNA, it is necessary to annotate the sequence of non-coding RNA, including finding the start and end sites of non-coding RNA, and then predicting its secondary structure and functional region; (5) Verification of regulatory RNA function: Verify the regulatory effect of non-coding RNA on polymerase I transcriptional activity in cell models and animal models, evaluate its impact on cell proliferation, differentiation and disease status, and evaluate the inhibitory effect of polymerase I on cancer cell proliferation in cell models; (6) RNA expression profiling: A large amount of non-coding RNA expression data can be obtained through high-throughput sequencing. RNA data provides a series of expression profiling methods and tools to help researchers explore the potential functions and regulatory mechanisms of non-coding RNA; (7) Functional verification: In order to further verify the function and mechanism of non-coding RNA, laboratory verification is required. Researchers design appropriate experimental plans and analyze experimental results to further verify the function and mechanism of non-coding RNA; Furthermore, in step (2), a non-coding RNA targeting a specific transcription factor is designed to bind to the transcription factor, thereby affecting its interaction with polymerase I, thereby achieving the purpose of mutual synthesis of different non-coding RNAs.

[0016] Furthermore, in step (3), the polymerase I transcription activity can be collected by adding the designed regulatory RNA to the in vitro transcription system and detecting the change in the RNA transcription level, thereby achieving precise regulation of the polymerase I transcription activity.

[0017] Furthermore, the step (4) predicts the secondary structure and functional region of the non-coding RNA through annotation of the non-coding RNA. The non-coding RNA data provides annotation tools and databases to help researchers annotate non-coding RNA sequences.

[0018] Experimental method: The experimental group adopted the method in comparative example 1, and the control group adopted the method in comparative example 2 (excluding the polymerase I association analysis step).

[0019] Criteria for judging the mechanism of regulating RNA polymerase I transcription: transcriptional activity, cell proliferation, cell differentiation control, and function of the regulatory mechanism.

[0020] The present invention can regulate protein conformation by regulating RNA function verification, RNA expression spectrum analysis and polymerase I association analysis, thereby adjusting the size of nucleolar proteins, and further affecting the transcriptional activity of RNA polymerase I. When the nucleolar proteins become smaller, the function of RNA polymerase I is hindered. Conversely, when the nucleolar proteins become larger, the inhibition of RNA polymerase I by nucleolar proteins can be released, and its efficient transcription can be promoted, so that non-coding RNA can regulate the interaction between RNA polymerase I and cells. Therefore, regulating the generation of rRNA is crucial to maintaining the normal function of cells. The regulatory effect of non-coding RNA on the transcriptional activity of polymerase I is verified in cell models and animal models, and its effects on cell proliferation, differentiation and disease status are evaluated, so as to enhance the inhibitory effect of polymerase I on the proliferation of cancer cells.

[0021] In the description of the present invention, it should be understood that the terms "longitudinal", "lateral", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside" and the like indicate positional relationships, which are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present invention. In the description of the present invention, unless otherwise specified and limited, it should be noted that the terms "installed", "connected" and "connected" should be understood in a broad sense, for example, it can be a mechanical connection or an electrical connection, or it can be the internal communication of two elements, it can be a direct connection, or it can be an indirect connection through an intermediate medium. For ordinary technicians in this field, the specific meanings of the above terms can be understood according to specific circumstances.

[0022] The above are only preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for studying the mechanism of non-coding RNA regulating RNA polymerase I transcription and its application, characterized in that: The steps of the RNA polymerase I transcription mechanism are as follows: (1) Collecting non-coding RNA data: In non-coding RNA research, it is necessary to collect databases and public data sets related to non-coding RNA, including the sequence, structure, function of non-coding RNA and transcriptomics data that integrates RNA expression levels. Some statistical methods and algorithms are used to reduce the dimension and standardize the data for subsequent analysis. (2) RNA data cleaning, design and synthesis: Based on the polymerase I transcription mechanism and the known RNA functions, RNA sequences with regulatory potential are designed, and the designed RNA molecules are obtained through chemical synthesis or biosynthesis. In order to better study the functions and regulatory mechanisms of non-coding RNA, data from different sources need to be integrated and analyzed; (3) Transcriptional regulatory mechanism: Using the molecular mechanism of the interaction between non-coding RNA data and polymerase I and its auxiliary factors, determine the key sites and signaling pathways of RNA data regulating polymerase I transcription, using in vitro transcription systems or cell models; (4) Annotation analysis of non-coding RNA: Annotation analysis is the process of matching unlabeled sequences with known information. In the study of non-coding RNA, it is necessary to annotate the sequence of non-coding RNA, including finding the start and end sites of non-coding RNA, and then predicting its secondary structure and functional region; (5) Verification of regulatory RNA function: Verify the regulatory effect of non-coding RNA on polymerase I transcriptional activity in cell models and animal models, evaluate its impact on cell proliferation, differentiation and disease status, and evaluate the inhibitory effect of polymerase I on cancer cell proliferation in cell models; (6) RNA expression profiling: A large amount of non-coding RNA expression data can be obtained through high-throughput sequencing. RNA data provides a series of expression profiling methods and tools to help researchers explore the potential functions and regulatory mechanisms of non-coding RNA; (7) Polymerase I association analysis: RNA analysis can be used to analyze the association between non-coding RNA and diseases. By integrating the disease database and the non-coding RNA database, the probability of association between non-coding RNA and diseases can be predicted, and correlation analysis and network analysis can be performed; (8) Functional verification: In order to further verify the function and mechanism of non-coding RNA, laboratory verification is required. Researchers design appropriate experimental plans and analyze the experimental results to further verify the function and mechanism of non-coding RNA.

2. A method for studying the mechanism of non-coding RNA regulating RNA polymerase I transcription and its application according to claim 1, characterized in that: The step (2) designs a non-coding RNA targeting a specific transcription factor, binds to the transcription factor, affects its interaction with polymerase I, and achieves the purpose of mutual synthesis of different non-coding RNAs.

3. The method for studying the mechanism of non-coding RNA regulating RNA polymerase I transcription and its application according to claim 1, characterized in that: In the step (3), the designed regulatory RNA is added to the in vitro transcription system to detect changes in the RNA transcription level, thereby collecting polymerase I transcription activity and achieving precise regulation of polymerase I transcription activity.

4. The method for studying the mechanism of non-coding RNA regulating RNA polymerase I transcription and its application according to claim 1, characterized in that: The step (4) predicts the secondary structure and functional region of non-coding RNA through annotation of non-coding RNA. Non-coding RNA data provides annotation tools and databases to help researchers annotate non-coding RNA sequences.

5. The method for studying the mechanism of non-coding RNA regulating RNA polymerase I transcription and its application according to claim 1, characterized in that: The step (7) utilizes the fact that non-coding RNA plays an important regulatory role in the occurrence and development of diseases, and utilizes biological big data technology to analyze the relationship between non-coding RNA and diseases.