A method for constructing a drosophila m6a RNA methylation tumor model and application thereof

Abstract

The application discloses a method for constructing a fruit fly m6A RNA methylation tumor model and application thereof. The application uses fruit flies as model organisms, and constructs an m6A RNA methylation tumor model by specifically overexpressing m6A related genes in fruit fly eye imaginal discs through a MARCM system. The method ensures the accuracy and reliability of the model, and provides a reliable in vivo tumor model for studying the role of m6A in tumor occurrence and development. The application fills the blank of m6A methylation in fruit fly tumor research by using the high visibility and non-invasive characteristics of the fruit fly model. The model constructed by the application can more accurately simulate the role of m6A methylation in in vivo tumors by specifically expressing m6A related genes and tumor inducing factors, and provides an important in vivo experimental tool for studying the molecular mechanism of m6A in tumor biology.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biomedical research, and specifically relates to a method for constructing a fruit fly m6A RNA methylation tumor model and its application. The model aims to systematically screen and verify the key molecular mechanisms of m6A methylation in the occurrence and development of tumors through fruit fly in vivo experiments, providing an important experimental tool for in-depth understanding of the role of m6A methylation in tumor biology. BACKGROUND

[0002] Drosophila melanogaster, as a classic model organism, has been widely used in genetic and developmental biology research due to its unique biological advantages. Drosophila has a short life cycle, strong reproductive ability, simple genome, and easy operation, making it an ideal model for studying gene function and complex biological processes. In addition, many genes and signaling pathways in Drosophila are highly conserved with humans, which makes the biological findings obtained in Drosophila often have important reference value.

[0003] Drosophila also has unique advantages in constructing tumor models. Through gene editing techniques such as the MARCM (Mosaic Analysis with a Repressible Cell Marker) system, specific genes can be overexpressed or knocked out in specific tissues of Drosophila, simulating the occurrence and development of human tumors. For example, the Drosophila eye imaginal disc is a commonly used model system, and overexpression of oncogenes (Ras and Raf activity) in the eye imaginal disc can observe the formation and growth of tumors, providing an intuitive experimental platform for studying the molecular mechanisms of tumor occurrence. The scribble gene encodes a cell polarity protein involved in maintaining cell polarity and tissue structural integrity. Drosophila with scribble gene mutations exhibit cell polarity disorder and tissue structure destruction, similar to the loss of cell polarity in human tumors. Introducing mutations in the scribble gene in cells overexpressing oncogenes (Ras and Raf activity) can lead to loss of cell polarity, promoting tumor formation. This model not only simulates the growth process of tumors, but also allows direct observation of tumor size and morphology under a microscope, providing a highly visible and non-invasive research tool.

[0004] RNA methylation, especially N6-methyladenosine (m6A) modification, has been a hot topic in recent years. m6A modification widely participates in various biological processes by regulating RNA transcription, maturation, translation, degradation, and stability. In vitro studies have shown that m6A modification is closely related to the occurrence and development of various cancers. For example, abnormal expression of m6A modification enzymes (such as METTL3, METTL14) and demethylases (such as FTO, ALKBH5) can affect the proliferation, metabolism, immune response, and microenvironment formation of tumor cells. However, the limitations of in vitro studies lie in the difficulty to fully reflect the complex physiological and pathological environment in vivo, therefore, in vivo studies are particularly important.

[0005] In in vivo studies, the role of RNA methylation has also been gradually revealed. Through studies in mouse models, m6A modification has been shown to play a key role in the occurrence and development of tumors. For example, m6A modification can regulate the expression of tumor-related genes and affect the proliferation and metastasis of tumor cells. However, these studies are often limited by high cost, complex operation, and long experimental period, which makes Drosophila a more ideal model organism.

[0006] Although the study of RNA methylation in Drosophila is relatively less, some preliminary findings have been made. m6A RNA methyltransferase also exists in Drosophila and plays an important role in Drosophila development. The m6A methylation regulatory factors in Drosophila have high homology with their counterparts in mammals, which provides a unique advantage for studying m6A methylation using Drosophila models. For example, the m6A methyltransferase complex mettl3 / mettl14 / nito in Drosophila corresponds to METTL3 / METTL14 / RBM15 in mammals, and these proteins are highly similar in structure and function. By constructing a Drosophila m6A RNA methylation tumor model, the role of m6A modification in tumor occurrence and development can be more systematically studied.

[0007] However, the specific mechanisms and functions of Drosophila m6A RNA methylation in in vivo tumors are not yet clear. The existing mouse in vivo tumor model construction methods are complex and difficult to efficiently, quickly, and accurately simulate the role of m6A in tumor occurrence and development, which seriously hinders the in-depth study of m6A methylation in in vivo tumors. Therefore, the present invention aims to develop an efficient and accurate method for constructing a Drosophila m6A RNA methylation tumor model to overcome the shortcomings of the prior art. SUMMARY

[0008] The problem to be solved by the present application is how to construct an m6A RNA methylation tumor model in fruit flies: the present application uses fruit flies as model organisms to construct an m6A RNA methylation tumor model by specifically overexpressing m6A related gene nito (homolog of human m6A RNA methylation regulator RBM15) in the eye imaginal disc of fruit flies through the MARCM system. This method ensures the accuracy and reliability of the model, providing a reliable in vivo tumor model for studying the role of m6A in tumor occurrence and development. The present application takes advantage of the high visibility and non-invasive nature of fruit fly models to fill the gap in the study of m6A methylation in fruit fly tumors. After the model is constructed, high-throughput mutation screening and systematic functional verification experiments can be used to confirm the specific role of the screened genes in tumor occurrence and development, providing a new perspective and tool for in-depth understanding of the function of m6A in tumors. The model can more accurately simulate the role of m6A methylation in in vivo tumors by specifically expressing m6A related genes and tumor inducing factors, providing an important in vivo experimental tool for studying the molecular mechanisms of m6A in tumor biology.

[0009] The present application adopts the following technical solutions:

[0010] A method for constructing a fruit fly m6A RNA methylation tumor model, specifically comprising the following steps:

[0011] The ey-FLP1;actin>y + >Gal4,UAS-GFP;FRT82B,Tubulin-Gal80 fruit fly strain is crossed with UAS-nito;UAS-Raf GOF ,FRT82B,scrib - / - / SM6B-TM6B.Tb fruit fly strain; the fruit fly m6A RNA methylation tumor model is constructed in the eye imaginal disc, and the fruit fly with the genotype ey-FLP1;actin>y + >Gal4,UAS-GFP;UAS-nito;UAS-Raf GOF ,FRT82B,scrib - / - / FRT82B,scrib - / - is obtained.

[0012] The present application overexpresses Raf GOF in the eye imaginal disc cells of fruit flies by MARCM technology, and mutates the polarity related gene scribble, and simultaneously overexpresses the RNA methylase nito, thereby constructing an m6A RNA methylation high expression tumor model in the eye imaginal disc of fruit flies. Compared with the closest prior art, the biggest difference of the present application is that through specific gene operation and multi-step combination, the construction of the high-efficiency m6A RNA methylation tumor model is realized.

[0013] Firstly, the present application overexpresses the Raf GOF gene in Drosophila eye imaginal disc cells through the MARCM system GOF Overexpression of the Raf GOF gene is a key step in constructing a tumor model, and through this operation, the formation of tumors in Drosophila eye imaginal discs can be induced. This technical means ensures the reliability and stability of the tumor model.

[0014] Secondly, the present application enhances tumor formation by mutating the polarity-related gene scribble. Mutation of the scribble gene leads to loss of cell polarity, further promoting tumor growth. This step not only increases the complexity and authenticity of the tumor model, but also provides a new perspective for studying the role of cell polarity in tumor development.

[0015] Finally, the present application overexpresses RNA methylase nito on the basis of the above. Overexpression of nito can significantly increase the level of m6A RNA methylation in Drosophila eye imaginal discs, and the tumor model with high expression of m6A RNA has stronger proliferation and invasion ability. This technical means is the core innovation of the present application, through the synergistic operation of multiple genes, not only a high-efficiency m6A RNA methylation tumor model is successfully constructed, but also a new tool for studying the role of m6A methylation in tumor development is provided.

[0016] In summary, the biggest difference of the present application is to increase and change multiple key steps in the manufacturing method, including specific overexpression of the Raf GOF gene, mutation of the scribble gene, and overexpression of the RNA methylase nito, which realizes the construction of a high-efficiency m6A RNA methylation tumor model. These specific gene operations and multi-step combinations not only ensure the accuracy and reliability of the model, but also fill the gap of m6A methylation in Drosophila tumor research.

[0017] The invention principle of the present application is:

[0018] The MARCM system is used to specifically overexpress m6A-related genes (such as METTL3, METTL14, WTAP, etc.) and the key kinase Raf in the Ras / MAPK signaling pathway in the adult eye disc of Drosophila, and then the FLP / FRT system is used to introduce homozygous mutations of the polarity-related gene scribble during cell mitosis, thereby constructing a Drosophila in vivo tumor model with high expression of m6A RNA. Overexpression of m6A-related genes can promote the proliferation and growth of tumor cells, thereby forming an obvious tumor phenotype. The advantages of Drosophila, such as high visibility, easy operation and low cost, can more efficiently screen and verify key factors regulating m6A methylation, provide a new perspective and tool for in-depth understanding of the function of m6A in tumors, and fill the gap of m6A methylation in Drosophila tumor research.

[0019] The beneficial effects of the present application are:

[0020] The present application first uses the MARCM system to specifically overexpress m6A-related genes in the adult eye disc of Drosophila, and successfully constructs a Drosophila tumor model with high expression of m6A RNA methylation. This precise gene regulation method ensures the accuracy and reliability of the model, providing a reliable platform for studying the role of m6A in tumor development.

[0021] The Drosophila m6A RNA methylation tumor model constructed in the present application can be used to verify potential drug targets and accelerate the early stage of drug discovery. The model can be used to evaluate the effect of candidate drugs in vivo, improve the success rate and efficiency of drug development. By screening and verifying m6A methylation-related genes, new biomarkers can be found for early diagnosis and prognosis evaluation of tumors. Understanding the specific role of m6A methylation in tumors helps to develop personalized treatment plans and improve treatment effectiveness. BRIEF DESCRIPTION OF DRAWINGS

[0022] Figure 1 The figure is a schematic diagram of the MARCM system. The system combines the FLP / FRT recombination system and the UAS / Gal4 system, uses FLP recombinase to cut the chromosomal fragments between FRT sites at a specific time point, removes the Gal80 inhibitor, and enables the Gal4-driven UAS gene to be expressed in specific cells, thereby achieving precise regulation and labeling of target genes.

[0023] Figure 2 UAS-nito; UAS-Raf GOF ,FRT82B,scrib - / - Flow chart of construction of Tb Drosophila strain.

[0024] Figure 3A flowchart for constructing m6A methylated tumors in Drosophila. The MARCM system is used to simultaneously express activated Raf, nito, and mutated scribble genes in cells. During mitosis, chromosomal recombination occurs under the action of FLP / FRT elements, resulting in three genotypes of cells.

[0025] Figure 4 A flowchart for constructing m6A methylated tumors in Drosophila. The m6A methylated tumor model is constructed in the Drosophila adult eye disc using the MARCM system. The GFP cells represent tumor cells, and the larger the GFP area, the stronger the tumor proliferation ability. DETAILED DESCRIPTION

[0026] This application uses Drosophila genetic crossing and recombination methods to overexpress Raf GOF and mutate scribble in the Drosophila adult eye disc using the Gal4 / UAS system, MARCM system, and overexpress RNA methylase nito, thereby inducing m6A RNA methylation high-expression tumors in the adult eye disc. This model can be used to study the role of m6A methylation in tumor development and is a novel in vivo tumor model for studying m6A methylation regulation signals.

[0027] The MARCM system is a powerful tool for cell-specific gene manipulation in Drosophila. This system combines the FLP / FRT recombination system and the UAS / Gal4 system to activate or inhibit specific genes in specific cells. Specifically, the MARCM system uses FLP recombinase to cut the chromosomal fragments between FRT sites at a specific time point, thereby removing the Gal80 inhibitor, allowing the UAS gene driven by Gal4 to be expressed. In this way, precise regulation of target genes in specific cells or tissues can be achieved (such as Figure 1 ).

[0028] N6-methyladenosine (m6A) is one of the most common modifications in RNA, which widely participates in various biological processes by regulating RNA transcription, maturation, translation, degradation, and stability. Nito is an RNA methylase, a homolog of human RBM15, which can catalyze the formation of m6A modification. In Drosophila, overexpression of nito can significantly increase RNA methylation levels, thereby affecting gene expression and cell function. Therefore, by overexpressing nito in Drosophila, a model of high expression of m6A RNA methylation can be constructed to study the role of m6A modification in tumor development.

[0029] Fruit flies are usually reared in standard fruit fly tubes or vials, with a cornmeal medium at the bottom and the tube or vial plugged with a latex sponge plug to ensure aeration. The medium is prepared by mixing cornmeal, yeast, sugar and water, and sometimes a small amount of acetic acid is added to prevent mold growth. Fruit flies grow best in a constant temperature environment of about 25°C, and the life cycle of each generation is about 10-14 days.

[0030] To construct an efficient m6A RNA methylation tumor model, we use two key fruit fly strains and achieve precise regulation of genes by crossing. Fruit flies have four chromosomes, and the present application mainly crosses and recombines on chromosomes 2, 3 and X.

[0031] The tool fruit flies needed include:

[0032] a. ey-FLP1; Sp / CyO; Sb / TM6B.Tb: Fruit flies carrying the recombinase FLP1 specifically expressed in the eye disc on the X chromosome, and balancers on chromosomes 2 and 3, with armpit hair, rolled wings, shoulder hair and truncated hair.

[0033] b. FRT82B, located on chromosome 3, white eye color, carrying a neo-resistance gene, a tool fruit fly for recombination.

[0034] c. Sp / CyO; Sb / TM6B.Tb and sco / CyO; Sb / TM6B.Tb are tool fruit flies for balancing.

[0035] d. Sp; Sb / SM6B-TM6B.Tb is a genetically coupled tool fruit fly, which carries balancers on chromosomes 2 and 3, respectively, showing armpit hair (Sb), rolled wings (CyO) and shoulder hair and truncated hair (TM6B.Tb). The two balancer chromosomes SM6B-TM6B.Tb are linked together and inherited, ensuring the stability of the genotype during crossing.

[0036] In this project we need to construct the following two genotypes of fruit flies for experiments:

[0037] 1. Fruit fly strain 1: ey-FLP1; actin>y + >Gal4, UAS-GFP; FRT82B, Tubulin-Gal80

[0038] ey-FLP1 (X chromosome): ey-FLP1 is a strain that specifically expresses FLP recombinase in the eye disc, which is used to initiate FLP / FRT recombination during mitosis.

[0039] actin>y +> Gal4 (2nd chromosome): This is a generic Gal4 driver system to activate the expression of UAS genes in specific cells.

[0040] UAS-GFP (2nd chromosome): UAS-GFP is a reporter gene to mark cells expressing Gal4 for easy observation and screening.

[0041] FRT82B, Tubulin-Gal80 (3rd chromosome): Tubulin-Gal80 is a generic Gal80 suppressor to suppress the expression of Gal4-driven UAS genes before FLP recombination.

[0042] The method for constructing the Drosophila strain 1 is as follows:

[0043] First, the actin>y + > Gal4 Drosophila strain is crossed with the UAS-GFP Drosophila strain to obtain the actin>y + > Gal4 / UAS-GFP female virgin Drosophila is crossed with the Sp / CyO; Sb / TM6B.Tb male Drosophila for two generations to obtain the actin>y + > Gal4, UAS-GFP / CyO; Sb / TM6B.Tb.

[0044] Subsequently, the FRT82B Drosophila strain is crossed with the Tubulin-Gal80 Drosophila strain to obtain the FRT82B / Tubulin-Gal80 female virgin Drosophila, which is then crossed with the Sp / CyO; Sb / TM6B.Tb male Drosophila to obtain the Sp / CyO; FRT82B, Tubulin-Gal80 / TM6B.Tb Drosophila strain.

[0045] Then, the actin>y + > Gal4, UAS-GFP / CyO; Sb / TM6B.Tb Drosophila strain is crossed with the Sp / CyO; FRT82B, Tubulin-Gal80 / TM6B.Tb Drosophila strain to construct the actin>y + > Gal4, UAS-GFP / CyO; FRT82B, Tubulin-Gal80 / TM6B.Tb Drosophila strain.

[0046] Finally, the ey-FLP1; Sp / CyO; Sb / TM6B.Tb female and male virgin Drosophila is crossed with the actin>y +Gal4, UAS-GFP / CyO; FRT82B, Tubulin-Gal80 / TM6B.Tb male flies were crossed to obtain ey-FLP1; actin>y+>Gal4, UAS-GFP; FRT82B, Tubulin-Gal80 female flies.

[0047] 2. Drosophila strain 2: UAS-nito; UAS-Raf GOF FRT82B, scrib - / - / SM6B-TM6B.Tb

[0048] UAS-nito (2nd chromosome): UAS-nito is a strain that expresses nito gene under the control of Gal4, which is used to overexpress nito in specific cells.

[0049] UAS-Raf GOF (3rd chromosome): UAS-Raf is a strain that expresses Raf activation fragment under the control of Gal4. GOF

[0050] FRT82B, scrib - / - (3rd chromosome): scrib is a strain that mutates the polarity-related gene scribble on the 3rd chromosome of Drosophila, which is used to simulate the loss of cell polarity. - / -

[0051] The construction process of Drosophila strain 2 is as follows Figure 2 , and the specific method is as follows:

[0052] First, UAS-Raf GOF FRT82B, scrib - / - Drosophila strain is crossed with sco / CyO; Sb / TM6B.Tb Drosophila strain for two generations to obtain sco / CyO; UAS-Raf GOF FRT82B, scrib - / - / TM6B.Tb fruit fly. UAS-nito is crossed with sco / CyO; Sb / TM6B.Tb Drosophila strain for two generations to obtain UAS-nito / CyO; Sb / TM6B.Tb Drosophila strain.

[0053] Then, sco / CyO; UAS-Raf GOF FRT82B, scrib - / - / TM6B.Tb Drosophila strain is crossed with UAS-nito / CyO; Sb / TM6B.Tb Drosophila strain to obtain UAS-nito / CyO; UAS-Raf GOF ​​FRT82B, scrib - / - / TM6B.Tb fruit fly strain, and then cross with Sp; Sb / SM6B-TM6B.Tb, select UAS-nito; UAS-Raf GOF FRT82B, scrib - / - / SM6B-TM6B.Tb genotype fruit fly.

[0054] By crossing the above two fruit fly strains, precise gene regulation and tumor model construction can be achieved.

[0055] 1. Cross process:

[0056] Cross fruit fly strain 1 virgin fly (ey-FLP1; actin>y + >Gal4, UAS-GFP; FRT82B, Tubulin-Gal80) with fruit fly strain 2 male fly (UAS-nito; UAS-Raf GOF FRT82B, scrib - / - / SM6B-TM6B.Tb).

[0057] 2. Gene expression regulation:

[0058] In the eye imaginal disc, ey-FLP1 will initiate FLP recombination during the cell mitosis period, cutting off Tubulin-Gal80 between FRT82B sites, removing the Gal80 inhibitor, allowing Gal4 to be freely expressed, and the scribble gene is homozygous mutation.

[0059] The Gal4-driven UAS system will activate the expression of UAS-Raf GOF , UAS-nito and UAS-GFP, respectively, to achieve overexpression of Raf, overexpression of nito and expression of GFP.

[0060] 3. Specific genotype ( Figure 3 ):

[0061] Through crossing, the genotype of the offspring fruit fly is: ey-FLP1; actin>y GOF >Gal4, UAS-GFP / UAS-nito; FRT82B, Tubulin-Gal80 / UAS-Raf - / - .

[0062] During mitosis, under the action of FLP / FRT elements, after chromosome recombination, three genotypes of cells will be formed, including:

[0063] a. ey-FLP1; actin>y+ >Gal4, UAS-GFP; UAS-nito; UAS-Raf GOF ,FRT82B,scrib - / - / FRT82B,

[0064] scrib - / - The scribble in this cell is a pure mutation, and expresses active Raf and nito, and is labeled with GFP as green fluorescence.

[0065] b.ey-FLP1; actin>y + >Gal4, UAS-GFP; UAS-nito; UAS-Raf GOF ,FRT82B,Tubulin-Gal80 /

[0066] FRT82B, Tubulin-Gal80. In this cell, no other genes are expressed due to the presence of Tubulin-Gal80, and the cell does not fluoresce.

[0067] c.ey-FLP1; actin>y+>Gal4, UAS-GFP; UAS-nito; UAS-Raf GOF ,FRT82B,scrib - / - / FRT82B,

[0068] Tubulin-Gal80. In this cell, the scribble gene is a heterozygous mutation, and Tubulin-Gal80 is present, so other genes are not expressed, and the cell does not fluoresce.

[0069] Through the above hybridization and gene expression regulation, a tumor model with high expression of m6A RNA methylation in the adult eye disc of Drosophila was successfully constructed. The genotype of the Drosophila selected as ey-FLP1; actin>y + >Gal4, UAS-GFP; UAS-nito; UAS-Raf GOF ,FRT82B,scrib - / - / FRT82B,scrib - / - The Drosophila m6A RNA methylation tumor model can be obtained. The effectiveness of the model is verified by observing and analyzing the size and morphology of the tumor.

[0070] In this model, overexpression of active Raf and nito, and mutation of scribble increase the level of m6A RNA methylation, and promote the proliferation and invasion ability of the tumor (such as Figure 4). By labeling tumor cells with GFP, it is easy to visually detect and analyze the size and morphology of tumors, and to screen and verify key factors that regulate m6A methylation, providing a powerful tool for in-depth study of the role of m6A in tumor occurrence and development.

Claims

1. A method for constructing a Drosophila m6A RNA methylation tumor model, characterized in that, The construction method specifically comprises the following steps: The ey-FLPl; actin > y + > Gal4, UAS-GFP; FRT82B, Tubulin-Gal80 fruit fly strain is crossed with UAS-nito; UAS-Raf GOF , FRT82B, scrib - / - / SM6B-TM6B.Tb fruit fly strain; construct the fruit fly m6A RNA methylation tumor model in the eye imaginal disc, and select the genotype of ey-FLPl; actin > y + > Gal4, UAS-GFP;UAS-nito; UAS-Raf GOF , FRT82B, scrib - / - / FRT82B, scrib - / - fruit fly The fruit fly m6A RNA methylation tumor model is constructed in the eye imaginal disc, and the specific method is as follows: In the eye imaginal disc, ey-FLP initiates FLP recombination at the cell mitotic stage, cuts off Tubulin-Gal80 between FRT82B sites, removes the Gal80 inhibitor, makes Gal4 freely express, and the scribble gene is homozygous mutation; The Gal4-driven UAS system activates the expression of UAS-Raf GOF , UAS-nito and UAS-GFP, which results in overexpression of Raf, overexpression of nito and expression of GFP, respectively.

2. The method for constructing a Drosophila m6A RNA methylation tumor model according to claim 1, characterized in that, The ey-FLP1; actin > y + The method for constructing the Drosophila strain of Gal4, UAS-GFP; FRT82B, Tubulin-Gal80 is as follows: the Drosophila strain of actin > y + The Drosophila strain of Gal4, UAS-GFP / CyO; Sb / TM6B.Tb is crossed with the Drosophila strain of Sp / CyO; FRT82B, Tubulin-Gal80 / TM6B.Tb, and the genotype of the selected Drosophila strain is actin > y + The Drosophila strain of Gal4, UAS-GFP / CyO; FRT82B, Tubulin-Gal80 / TM6B.Tb is crossed with the Drosophila strain of ey-FLP1; Sp / CyO; Sb / TM6B.Tb, and the Drosophila strain of ey-FLP1; actin > y is obtained + The Drosophila strain of Gal4, UAS-GFP; FRT82B, Tubulin-Gal80.

3. The method of claim 1, wherein the fruit fly m6A RNA methylation tumor model is constructed by, UAS-nito; UAS-Raf GOF , FRT82B, scrib - / - The method for constructing the D. melanogaster strain of sco / CyO; UAS-Raf GOF ,FRT82B, scrib - / - / TM6B.Tb is to cross the D. melanogaster of sco / CyO; Sb / TM6B.Tb with the D. melanogaster of UAS-Raf GOF ,FRT82B, scrib - / - / TM6B.Tb for two generations to construct the D. melanogaster strain of sco / CyO; UAS-Raf UAS-nito / CyO; Sb / TM6B.Tb flies were constructed by crossing UAS-nito with sco / CyO; Sb / TM6B.Tb flies for two generations; then, sco / CyO; UAS-Raf GOF ,FRT82B, scrib - / - / TM6B.Tb flies were crossed with UAS-nito / CyO; Sb / TM6B.Tb flies to obtain UAS-nito / CyO; UAS-Raf GOF ,FRT82B, scrib - / - / TM6B.Tb flies, and then crossed with Sp; Sb / SM6B-TM6B.Tb to obtain UAS-nito; UAS-Raf GOF , FRT82B, scrib - / - / SM6B-TM6B.Tb flies.

4. Use of a Drosophila m6A RNA methylation tumor model, characterized in that, The fruit fly m6A RNA methylation tumor model is constructed by the method in any one of claims 1-3, and the model is used for discovering new tumor biomarkers and verifying potential tumor drug targets.

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