CN110567906A - A method and application for characterizing RNA methylation modification - Google Patents

Abstract

The present invention provides a method and application for characterizing RNA methylation modification. The method includes placing an RNA sample in a quartz plate, and using ultraviolet absorption spectroscopy and/or fluorescence spectroscopy to detect characteristic peaks of the RNA sample. The invention aims at the m6A methylation modification pathway of RNA, and applies the theoretical calculation method to obtain the characteristic absorption spectrum of the RNA sequence and the RNA m6A methylation sequence, and establishes a characterization method for the dynamic modification of RNA methylation based on the spectroscopy method. The migration of characteristic absorption peaks realizes the identification of mRNA m6A methylation/demethylation, and at the same time realizes the variable temperature quantitative analysis of RNA methylation modification under the action of RNA methylation modification enzymes Mettl3/Mettl14/WTAP. The method has application prospects in evaluating and screening inhibitors of RNA methylation modifying enzymes and the development of therapeutic drugs for related diseases.

Description

A method and application for characterizing RNA methylation modification

technical field

The invention belongs to the technical field of epitranscriptomics, and relates to a method and application for characterizing RNA methylation modification.

Background technique

In recent years, the reversibly methylated RNA N6-methyladenosine (m6A) has received more and more attention, and researchers have successively localized m6A in mammalian transcriptomes and identified the implementation of this dynamic modification. The functions of "reading", "writing" and "erasing" were analyzed, and the function of m6A in post-transcriptional regulation was gradually analyzed. The study found that in the epitranscriptome, m6A is the most common and abundant mRNA molecular modification in eukaryotes, and the functional study of m6A has expanded human understanding of the disease process. Mettl3/Mettl14/WTAP methyltransferase complex catalyzes m6A modification, FTO and ALKBH5 demethylase demethylates m6A, m6A modification enzymes and binding proteins YTHDF1, YTHDF2 and YTHDC1 are used to recognize m6A, demonstrating that RNA Methylation modification is a dynamically reversible process, and this dynamically reversible epitranscriptome plays an important role in a variety of biological processes. Not only does this process establish a normal cellular phenotype, but it can also lead to disease. For example, Kharas MG et al found that in human hematopoietic stem/progenitor cells (HSPC), m6A-modifying enzyme mettl3, which is depleted by shRNA-mediated depletion, can promote cell differentiation and reduce cell proliferation; in contrast, overexpression of wild-type, which has no catalytic activity, mettl3, can inhibit cell differentiation and promote cell growth. With the in-depth study of the function of m6A in post-transcriptional regulation, the dynamic modification process of RNA methylation and demethylation is closely related to the occurrence and development of various diseases.

The dynamic nature of epigenetic regulation means that when directly amplifying the molecular factors of disease-related processes, it is possible to alter disease-related epigenetic states. RNA methylation modification is dynamic and reversible, and it has far-reaching significance for drug development based on m6A regulatory proteins and clinical treatment and translation applications based on m6A regulation. In both basic and applied research on the relationship between m6A levels and diseases, efficient and sensitive quantitative detection of the dynamic levels of m6A methylation modification is required. At present, the research on m6A modification mainly focuses on function, at the molecular level, cellular level and in vivo animal (mostly mouse or rat) level, usually using PCR amplification, Western-blot, gene knockout or mutation, molecular Cell biology and molecular biology methods such as recombinant expression are cumbersome and difficult to operate when identifying RNA methylation and demethylation.

CN 105018617A discloses a single gene mRNA methylation level detection method, comprising the following steps: 1), designing fluorescent quantitative primers for target gene methylation regions; 2), extracting total RNA, and using chemical fragmentation method to fragment the total RNA into RNA fragments; 3), the fragmented RNA obtained in step 2) was precipitated by an antibody that specifically recognizes m6A, and all mRNA fragments with m6A sites were precipitated, and collected and enriched with agarose magnetic beads; 4), the The mRNA fragments obtained by antibody enrichment in step 3) are reverse transcribed, and the quantitative PCR primers of step 1) are used to perform real-time quantitative PCR to obtain the relative expression of target genes containing methylation. Represents the level of mRNA methylation of a single gene. The method is based on fluorescence quantitative PCR, and the process is cumbersome.

CN 106047997 A discloses a high-throughput detection method for mRNA methylation, comprising the following steps: extracting mRNA samples; fragmenting the extracted RNA samples; using the obtained RNA fragments to specifically identify m6A methylation (m6A) on mRNA The mRNA fragments containing m6A were eluted and purified by precipitation; the purified RNA was reverse transcribed into cDNA, and DNA end repair, adapter ligation and PCR amplification were performed to complete the mRNA Construction of a methylation library; the completed library is sequenced using a high-throughput sequencing platform that analyzes sequencing libraries. The method goes through multiple steps such as immune enrichment, precipitation purification, reverse transcription and PCR amplification, and the operation is complicated.

At present, the efficient and sensitive method applied to RNA epigenetic modification is still blank at home and abroad.

SUMMARY OF THE INVENTION

In view of the deficiencies and actual needs of the prior art, the present invention provides a method and application for characterizing RNA methylation modification. The method is based on a spectral analysis method and realizes mRNA m6A methylation/demethylation through the migration of characteristic absorption peaks. The identification of methylation, and the variable temperature quantitative analysis of RNA methylation modification can be realized, with good accuracy and sensitivity.

For this purpose, the present invention adopts the following technical solutions:

In a first aspect, the present invention provides a method for characterizing RNA methylation modification, the method comprising placing an RNA sample in a quartz plate, and using ultraviolet absorption spectroscopy and/or fluorescence spectroscopy to detect characteristic peaks of the RNA sample .

In the present invention, aiming at the m6A methylation modification pathway of RNA, the theoretical calculation method is used to obtain the characteristic absorption spectrum of the RNA sequence and the RNAm6A methylation sequence, and the characterization method of the dynamic modification of RNA methylation based on the spectroscopic method is established. , in the process of methylation modification of RNA, the ultraviolet absorption spectrum is red-shifted, and the emission spectrum intensity is enhanced. During the demethylation process of methylated RNA, the ultraviolet absorption spectrum is blue-shifted, and the emission spectrum intensity is weakened. The method realizes the identification of mRNA m6A methylation/demethylation through the migration of characteristic absorption peaks, and at the same time realizes the quantitative analysis of RNA methylation modification with temperature change. good accuracy and sensitivity.

Preferably, the detection wavelength of the ultraviolet absorption spectroscopy is 200-900 nm, for example, it can be 200 nm, 300 nm, 400 nm, 500 nm, 600 nm, 700 nm, 800 nm or 900 nm.

Preferably, the detection temperature of the ultraviolet absorption spectrometry is 25 to 50°C, for example, 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C , 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, 46°C, 47°C, 48°C, 49°C or 50°C, preferably It is 27～45 ℃.

Preferably, the detection pressure of the ultraviolet absorption spectrometry is standard atmospheric pressure.

Preferably, the detection wavelength of the fluorescence spectroscopy is 280-600 nm, such as 280 nm, 300 nm, 320 nm, 350 nm, 380 nm, 400 nm, 420 nm, 450 nm, 480 nm, 500 nm, 520 nm, 550 nm, 580 nm or 600 nm.

Preferably, the excitation light wavelength of the fluorescence spectroscopy is 250-260 nm, such as 250 nm, 251 nm, 252 nm, 253 nm, 254 nm, 255 nm, 256 nm, 257 nm, 258 nm, 259 nm or 260 nm.

Preferably, the detection temperature of the fluorescence spectrometry is 25 to 50°C, such as 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, 46°C, 47°C, 48°C, 49°C or 50°C, preferably 27～45℃.

Preferably, the detection time of the fluorescence spectroscopy is 10-30 μs, for example, 10 μs, 11 μs, 12 μs, 13 μs, 14 μs, 15 μs, 16 μs, 17 μs, 18 μs, 19 μs, 20 μs, 21 μs, 22 μs, 23 μs, 24 μs, 25 μs , 26μs, 27μs, 28μs, 29μs or 30μs.

Preferably, the detection pressure of the fluorescence spectrometry is standard atmospheric pressure.

Preferably, the RNA sample includes any one or a combination of at least two of single-stranded RNA, single-stranded methylated RNA, double-stranded RNA, double-stranded methylated RNA or tissue RNA.

In a second aspect, the present invention provides the use of spectroscopic methods in characterizing RNA methylation modifications, the spectroscopic methods including ultraviolet absorption spectroscopy and/or fluorescence spectroscopy.

Preferably, the detection wavelength of the ultraviolet absorption spectroscopy is 200-900 nm, for example, it can be 200 nm, 300 nm, 400 nm, 500 nm, 600 nm, 700 nm, 800 nm or 900 nm.

Preferably, the detection temperature of the ultraviolet absorption spectrometry is 25 to 50°C, for example, 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C , 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, 46°C, 47°C, 48°C, 49°C or 50°C, preferably It is 27～45 ℃.

Preferably, the detection pressure of the ultraviolet absorption spectrometry is standard atmospheric pressure.

Preferably, the detection wavelength of the fluorescence spectroscopy is 280-600 nm, such as 280 nm, 300 nm, 320 nm, 350 nm, 380 nm, 400 nm, 420 nm, 450 nm, 480 nm, 500 nm, 520 nm, 550 nm, 580 nm or 600 nm.

Preferably, the excitation light wavelength of the fluorescence spectroscopy is 250-260 nm, such as 250 nm, 251 nm, 252 nm, 253 nm, 254 nm, 255 nm, 256 nm, 257 nm, 258 nm, 259 nm or 260 nm.

Preferably, the detection temperature of the fluorescence spectrometry is 25 to 50°C, such as 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, 46°C, 47°C, 48°C, 49°C or 50°C, preferably 27～45℃.

Preferably, the detection time of the fluorescence spectroscopy is 10-30 μs, for example, 10 μs, 11 μs, 12 μs, 13 μs, 14 μs, 15 μs, 16 μs, 17 μs, 18 μs, 19 μs, 20 μs, 21 μs, 22 μs, 23 μs, 24 μs, 25 μs , 26μs, 27μs, 28μs, 29μs or 30μs.

Preferably, the detection pressure of the fluorescence spectrometry is standard atmospheric pressure.

Preferably, the RNA sample includes any one or a combination of at least two of single-stranded RNA, single-stranded methylated RNA, double-stranded RNA, double-stranded methylated RNA or tissue RNA.

In a third aspect, the present invention provides an application of the method according to the first aspect in screening for inhibitors and/or activators of RNA methylation-modifying enzymes.

In a fourth aspect, the present invention provides an inhibitor of an RNA methylation modification enzyme, which is obtained by screening the method described in the first aspect.

In a fifth aspect, the present invention provides an activator of an RNA methylation modification enzyme, and the activator is screened by the method described in the first aspect.

In the sixth aspect, the present invention provides an application of the inhibitor as described in the fourth aspect and/or the activator as described in the fifth aspect in the preparation of a medicament for the diagnosis and/or treatment of diseases related to RNA methylation modification .

Compared with the prior art, the present invention has the following beneficial effects:

(1) The characterization method for dynamic modification of RNA methylation based on spectroscopy of the present invention realizes the identification of mRNA m6A methylation/demethylation through the migration of characteristic absorption peaks. Under the characteristic absorption peaks, the absorbance and RNA methylation The concentration of methylated or demethylated products is linear, the detection limit can reach 2 μmol/L, and the accuracy can reach 99%;

(2) The method of the present invention realizes the variable temperature quantitative detection of RNA methylation modification in the range of 25-50°C, has the advantages of simplicity, speed, accuracy, low detection limit, easy operation, etc. Or semi-quantitative, complex operation and other defects.

Description of drawings

Fig. 1 is the UV-Vis absorption spectrum of mRNA single-stranded sequence and mRNA methylation-modified single-stranded sequence;

Fig. 2 is the fluorescence spectrum of mRNA single-stranded sequence and mRNA methylation-modified single-stranded sequence;

Figure 3(A) shows the UV absorption spectra of mRNA single-stranded sequences at different temperatures, and Figure 3(B) shows the UV absorption spectra of mRNA methylation-modified single-stranded sequences at different temperatures;

Figure 4(A) shows the fluorescence spectra of mRNA single-stranded sequences at different temperatures, and Figure 4(B) shows the fluorescence spectra of mRNA methylation-modified single-stranded sequences at different temperatures.

Detailed ways

In order to further illustrate the technical means adopted by the present invention and its effects, the present invention will be further described below with reference to the embodiments and accompanying drawings. It should be understood that the specific embodiments described herein are only used to explain the present invention, but not to limit the present invention.

If no specific technique or condition is indicated in the examples, the technique or condition described in the literature in the field or the product specification is used. The reagents or instruments used without the manufacturer's indication are conventional products that can be purchased through regular channels.

Example 1 Sample Preparation

This embodiment selects 5 kinds of samples, and the specific preparation method is as follows:

Sample 1: Single-stranded RNA sample, sequence GGACU, purchased from GenScript; in the intercellular fume hood, add 500 μL of sterile water to the dry powder of the above sequence, dilute it to a 20 μM working solution, shake slightly at 25°C, After centrifugation at low speed, let stand for 1 min to obtain sample 1;

Sample 2: Single-stranded RNA methylation sample, the sequence is GGm6ACU, purchased from GenScript; in the intercellular fume hood, add 500 μL of sterile water to the dry powder of the above sequence, and dilute to a 20 μM working solution at 25°C Slightly shake, centrifuge at low speed and let stand for 1 min to obtain sample 2;

Sample 3: Double-stranded RNA sample, the sequence is (GGACU)/(CCUGA), purchased from GenScript; the above two RNA single-strands were dissolved in sterile water respectively, mixed according to the molar ratio of 1:1, and placed in the Incubate in a water bath at 85°C for 15min, slowly cool to 4°C and let stand for 1h, the two single strands complement each other to form double-stranded RNA, that is, sample 3;

Sample 4: Double-stranded RNA methylation sample, the sequence is (GGm6ACU)/(CCUGA), purchased from GenScript; the above two RNA single-strands were dissolved in phosphate buffer solution, according to the molar ratio of 1:1 After mixing, it was placed in a water bath at 85°C for 15 minutes, slowly cooled to 4°C and left for 1 hour, the two single strands were complementary to form double-stranded RNA, and sample 4 was obtained;

Sample 5: Mouse tissue RNA, extracted by Trizol method, the steps are as follows:

(1) Take the muscle tissue of wild-type mice (WT) and mutant mice with specific knockout of the ml3 gene, and crush them in a tissue crusher (JXFSTPRP-24 Shanghai Jingxin Industrial Development Co., Ltd.);

(2) Add 3mL Trizol by blowing to break the tissue, transfer it into a new EP tube, shake slowly for 20s, and let stand for 5min at room temperature;

(3) Add 900 μL of chloroform, shake vigorously for 15 s, stand at room temperature for 3 min, centrifuge at 12000 rpm for 15 min at 4°C, and transfer the upper aqueous phase to a new EP tube;

(4) Add equal volume of isopropanol to the water phase, shake at medium speed for 10s and mix, let stand for 10min at -20°C, centrifuge at 12000rmp for 10min at 4°C, discard the supernatant;

(5) Add 1.5mL of 70% ethanol, shake at medium speed for 10s to wash the precipitate, centrifuge at 12000rmp for 10min at 4°C, discard the supernatant, absorb the residual liquid, and air dry for 10min in a clean place;

(6) Add an appropriate amount of DEPC water to dissolve, detect the concentration of the extracted RNA, and store it in a -80°C refrigerator.

Example 2 Ultraviolet absorption spectroscopy analysis of single-stranded RNA methylation/demethylation modification

Take 10μL of working solution of sample 1 and sample 2, dilute with 90μL of water, add to 96-well quartz plate, measure the absorbance at 200-900nm wavelength, the detection temperature is 27℃, the detection pressure is 1 atmosphere, and the detection instrument is BioTek micro Spectrophotometer Cytation 1 to acquire the absorption spectra of sample 1 and sample 2.

As shown in Figure 1, A represents sample 1, the coordinates of the strongest absorption peak are (257, 0.650), m6A represents sample 2, and the coordinates of the strongest absorption peak are (258, 0.618), which is the same as that of sample 1 (single-stranded RNA ), the UV absorption spectrum of sample 2 (single-stranded methylated RNA) undergoes a 1-2 nm red shift, and the light intensity decreases by 0.3.

Example 3 Fluorescence spectroscopy analysis of single-stranded RNA methylation/demethylation modification

Take 10 μL of the working solution of sample 1 and sample 2, dilute with 90 μL of water, add it to a 96-well quartz plate, detect the absorbance at a wavelength of 280-600 nm, the emission spectrum is 255 nm, the detection temperature is 27 ° C, the detection time is 20 μs, and the air pressure is detected. The pressure is 1 atmosphere, the detection instrument is the TESCAN multi-function microplate reader INFINITE M PLEX, and the absorption spectra of sample 1 and sample 2 are obtained.

As shown in Figure 2, A represents sample 1, the coordinates of the strongest absorption peak are (510, 158), m6A represents sample 2, the coordinates of the strongest absorption peak are (512, 165), and the characteristic peaks of both appear. At around 510, compared with the characteristic peak of sample 1 (single-stranded RNA), the light intensity of the characteristic peak of sample 2 (single-stranded methylated RNA) is enhanced, and the characteristic peak is red-shifted by 2 nm.

Example 4 UV absorption spectra of RNA methylation/demethylation modification at different temperatures

Compared with Example 2, the detection temperature of ultraviolet absorption spectrum was 37°C or 45°C, and other conditions were the same as those of Example 2.

As shown in Figure 3(A), A represents sample 1. The abscissas of the strongest absorption peaks at the three temperatures are all 258 nm. As the temperature increases from 25 °C to 37 °C, the peak value increases from 0.395 to 0.4096. When the temperature is raised to 45°C, the peak value drops to 0.409;

As shown in Figure 3(B), m6A represents sample 2. The abscissas of the strongest absorption peaks at the three temperatures are all 257 nm. As the temperature increases from 25 °C to 37 °C, the peak value increases from 0.4035 to 0.4163. Raising the temperature to 45°C, the peak dropped to 0.4133.

Example 5 Fluorescence spectra of RNA methylation/demethylation modification at different temperatures

Compared with Example 3, the detection temperature of the fluorescence spectrum was 37°C or 45°C, and other conditions were the same as those of Example 3.

As shown in Figure 4(A), A represents sample 1. As the temperature increases from 25°C to 37°C and 45°C, the ordinate of the peak decreases from 172 to 159.3333 and 144.3333, respectively;

As shown in Figure 4(B), m6A represents sample 2. As the temperature increases from 25°C to 37°C and 45°C, the ordinate of the peak decreases from 154.3333 to 146.3333 and 138.667, respectively, indicating that with the increase of temperature, the RNA Distinction between UV-vis absorption and m6A methylated RNA was significantly improved.

Example 6 UV absorption spectroscopic analysis of double-stranded RNA methylation/demethylation modification

Take 10 μL of the working solution of sample 3 and sample 4, dilute with 90 μL of water, add it to a 96-well quartz plate, measure the absorbance at a wavelength of 200-900 nm, the detection temperature is 27 °C, and the detection pressure is 1 atmosphere. Spectrophotometer Cytation 1 to acquire the absorption spectra of sample 3 and sample 4.

Example 7 Fluorescence spectroscopic analysis of double-stranded RNA methylation/demethylation modification

Take 10 μL of the working solution of sample 3 and sample 4, dilute with 90 μL of water, add it to a 96-well quartz plate, detect the absorbance at a wavelength of 280-600 nm, the emission spectrum is 255 nm, the detection temperature is 27 ° C, the detection time is 20 μs, and the air pressure is detected. The pressure is 1 atmosphere, the detection instrument is the TESCAN multi-function microplate reader INFINITE M PLEX, and the absorption spectra of sample 3 and sample 4 are obtained.

Example 8 Ultraviolet absorption spectroscopy analysis of mouse tissue RNA methylation/demethylation modification

Take 10 μL of the extracted wild-type mouse muscle tissue RNA and mutant mouse muscle tissue RNA, add them to a 96-well quartz plate, detect the absorbance at a wavelength of 200-900nm, the detection temperature is 27 °C, and the detection pressure is 1 atmosphere. The detection instrument was BioTek microspectrophotometer Cytation 1, and the absorption spectrum of RNA methylation/demethylation modification was obtained.

Example 9 Fluorescence spectroscopy analysis of mouse tissue RNA methylation/demethylation modification

Take 10 μL of the extracted wild-type mouse muscle tissue RNA and mutant mouse muscle tissue RNA, add it to a 96-well quartz plate, detect the absorbance at a wavelength of 280-600nm, the emission spectrum is 255nm, the detection temperature is 27 ℃, and the detection time For 20 μs, the detection pressure is 1 atmosphere, the detection instrument is TESCAN multi-function microplate reader INFINITE M PLEX, and the absorption spectrum of RNA methylation/demethylation modification is obtained.

Example 10 Detection Limit Experiment

Take 10 μL of working solution of sample 1 and sample 2, dilute with 90 μL of water, and make 5-fold, 10-fold, 20-fold, 40-fold, 80-fold and 120-fold equal dilutions, respectively, and place the equally diluted sample and blank sample respectively. Measure the absorbance under the BioTek microspectrophotometer, use the concentration and absorbance as the standard curve, the linear part is the detectable concentration of UV-Vis absorption analysis, the lower end of the S-shaped curve is the lower limit of the detection range, and the detection limit is 2 μmol/L; The above samples were detected under the TESCAN multi-function microplate reader INFINITE M PLEX, and the concentration and luminosity were used as the standard curve to obtain the lowest detection limit, which was 2 μmol/L.

To sum up, the characterization method for dynamic modification of RNA methylation based on spectroscopy of the present invention realizes the identification of mRNA m6A methylation/demethylation through the migration of characteristic absorption peaks. The concentration of methylation products or demethylation products has a linear relationship, the detection limit can reach 20 μmol/L, and the accuracy can reach 99%. It has the advantages of simple, fast, accurate, low detection limit, easy operation, etc., and overcomes the defects of the existing technology, such as only qualitative or semi-quantitative, and complicated operation.

The applicant declares that the present invention illustrates the detailed method of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned detailed method, that is, it does not mean that the present invention must rely on the above-mentioned detailed method to be implemented. Those skilled in the art should understand that any improvement of the present invention, the equivalent replacement of each raw material of the product of the present invention, the addition of auxiliary components, the selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims (10)

1. A method for characterizing RNA methylation modification, wherein the method comprises placing the RNA sample in a quartz plate, and using ultraviolet absorption spectroscopy and/or fluorescence spectroscopy to detect the characteristic peaks of the RNA sample. 2. The method according to claim 1, wherein the detection wavelength of the ultraviolet absorption spectrometry is 200-900 nm; Preferably, the detection temperature of the ultraviolet absorption spectrometry is 25-50°C, preferably 27-45°C; Preferably, the detection pressure of the ultraviolet absorption spectrometry is standard atmospheric pressure; Preferably, the detection wavelength of the fluorescence spectroscopy is 280-600 nm; Preferably, the excitation light wavelength of the fluorescence spectroscopy is 250-260 nm; Preferably, the detection temperature of the fluorescence spectrometry is 25-50°C, preferably 27-45°C; Preferably, the detection time of the fluorescence spectroscopy is 10-30 μs; Preferably, the detection pressure of the fluorescence spectrometry is standard atmospheric pressure. 3. The method according to claim 1 or 2, wherein the RNA sample comprises any one of single-stranded RNA, single-stranded methylated RNA, double-stranded RNA, double-stranded methylated RNA or tissue RNA one or a combination of at least two. 4. The application of spectroscopy in characterizing RNA methylation modification, characterized in that, the spectroscopy comprises ultraviolet absorption spectroscopy and/or fluorescence spectroscopy. 5. The application according to claim 4, wherein the detection wavelength of the ultraviolet absorption spectrometry is 200-900 nm; Preferably, the detection temperature of the ultraviolet absorption spectrometry is 25-50°C, preferably 27-45°C; Preferably, the detection pressure of the ultraviolet absorption spectrometry is standard atmospheric pressure; Preferably, the detection wavelength of the fluorescence spectroscopy is 280-600 nm; Preferably, the excitation light wavelength of the fluorescence spectroscopy is 250-260 nm; Preferably, the detection temperature of the fluorescence spectrometry is 25-50°C, preferably 27-45°C; Preferably, the detection time of the fluorescence spectroscopy is 10-30 μs; Preferably, the detection pressure of the fluorescence spectrometry is standard atmospheric pressure. 6. The application according to claim 4 or 5, wherein the RNA sample comprises any one of single-stranded RNA, single-stranded methylated RNA, double-stranded RNA, double-stranded methylated RNA or tissue RNA one or a combination of at least two. 7. Application of the method according to any one of claims 1 to 3 in screening for inhibitors and/or activators of RNA methylation modification enzymes. 8. An inhibitor of RNA methylation modification enzyme, characterized in that, the inhibitor is obtained by screening the method according to any one of claims 1-3. 9. An activator of an RNA methylation modification enzyme, characterized in that, the activator is obtained by screening the method according to any one of claims 1-3. 10. Use of the inhibitor as claimed in claim 8 and/or the activator as claimed in claim 9 in the preparation of a medicament for diagnosis and/or treatment of diseases related to RNA methylation modification.