The miR-6240 target gene Igf2bp3 promotes myoblast fusion by enhancing myomaker mRNA stability
Abstract Background Myoblast fusion plays a crucial role in myogenesis. Insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3) functions as an RNA N 6-methyladenosine reader and exerts important roles in various biological processes. While our prior study suggested Igf2bp3 contributes to myog...
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| Format: | Article |
| Language: | English |
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BMC
2024-12-01
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| Series: | Cellular & Molecular Biology Letters |
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| Online Access: | https://doi.org/10.1186/s11658-024-00650-1 |
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| author | Yuxin Huang Wei Wang Xinhao Fan Xiaoqin Liu Weiwei Liu Zishuai Wang Yixing Li Yalan Yang Zhonglin Tang |
| author_facet | Yuxin Huang Wei Wang Xinhao Fan Xiaoqin Liu Weiwei Liu Zishuai Wang Yixing Li Yalan Yang Zhonglin Tang |
| author_sort | Yuxin Huang |
| collection | DOAJ |
| description | Abstract Background Myoblast fusion plays a crucial role in myogenesis. Insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3) functions as an RNA N 6-methyladenosine reader and exerts important roles in various biological processes. While our prior study suggested Igf2bp3 contributes to myogenesis, its molecular regulatory mechanism is largely unclear. Methods Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot were used for gene expression analysis. siRNA and CRISPRi technologies were conducted to knockdown the expression of Igf2bp3. CRISPR/Cas9 technology was performed to knockout Igf2bp3. The Igf2bp3 overexpression vector was designed using the pcDNA3.1(+) vector. Immunofluorescence detection was employed for subcellular localization and cell differentiation analysis. Cell Counting Kit-8 (CCK-8) and 5-ethynyl-2′-deoxyuridine (EdU) assays were conducted for cell proliferation and fusion detection. The dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were utilized for regulatory mechanism analysis of Igf2bp3. Results The overexpression of Igf2bp3 enhances myoblast fusion while knockdown of Igf2bp3 blocks the formation of myotubes. miR-6240 promotes myoblast proliferation while preventing myoblast differentiation and fusion by targeting the 3′ untranslated rgion (UTR) of Igf2bp3. Notably, the impacts of miR-6240 mimics on myoblast proliferation, differentiation, and fusion can be effectively counteracted by the overexpression of Igf2bp3. Moreover, our findings elucidate a direct interaction between Igf2bp3 and the myoblast fusion factor myomaker (Mymk). Igf2bp3 binds to Mymk to enhance its mRNA stability. This interaction results in increased expression of Mymk and heightened myoblast fusion. Conclusions Our study unveils Igf2bp3 as a novel post-transcriptional regulator of myoblast fusion through the miR-6240/Mymk axis, significantly contributing to our understanding of skeletal muscle development. Graphical Abstract |
| format | Article |
| id | doaj-art-4b6c8ab6bcca49cf8090d4bd6ab3195e |
| institution | Kabale University |
| issn | 1689-1392 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Cellular & Molecular Biology Letters |
| spelling | doaj-art-4b6c8ab6bcca49cf8090d4bd6ab3195e2024-12-08T12:37:24ZengBMCCellular & Molecular Biology Letters1689-13922024-12-0129112010.1186/s11658-024-00650-1The miR-6240 target gene Igf2bp3 promotes myoblast fusion by enhancing myomaker mRNA stabilityYuxin Huang0Wei Wang1Xinhao Fan2Xiaoqin Liu3Weiwei Liu4Zishuai Wang5Yixing Li6Yalan Yang7Zhonglin Tang8Kunpeng Institute of Modern Agriculture at Foshan, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural SciencesKunpeng Institute of Modern Agriculture at Foshan, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural SciencesKunpeng Institute of Modern Agriculture at Foshan, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural SciencesKunpeng Institute of Modern Agriculture at Foshan, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural SciencesKunpeng Institute of Modern Agriculture at Foshan, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural SciencesKunpeng Institute of Modern Agriculture at Foshan, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural SciencesGuangxi Key Laboratory of Animal Breeding, Disease Control and Prevention; College of Animal Science and Technology, Guangxi UniversityKunpeng Institute of Modern Agriculture at Foshan, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural SciencesKunpeng Institute of Modern Agriculture at Foshan, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural SciencesAbstract Background Myoblast fusion plays a crucial role in myogenesis. Insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3) functions as an RNA N 6-methyladenosine reader and exerts important roles in various biological processes. While our prior study suggested Igf2bp3 contributes to myogenesis, its molecular regulatory mechanism is largely unclear. Methods Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot were used for gene expression analysis. siRNA and CRISPRi technologies were conducted to knockdown the expression of Igf2bp3. CRISPR/Cas9 technology was performed to knockout Igf2bp3. The Igf2bp3 overexpression vector was designed using the pcDNA3.1(+) vector. Immunofluorescence detection was employed for subcellular localization and cell differentiation analysis. Cell Counting Kit-8 (CCK-8) and 5-ethynyl-2′-deoxyuridine (EdU) assays were conducted for cell proliferation and fusion detection. The dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were utilized for regulatory mechanism analysis of Igf2bp3. Results The overexpression of Igf2bp3 enhances myoblast fusion while knockdown of Igf2bp3 blocks the formation of myotubes. miR-6240 promotes myoblast proliferation while preventing myoblast differentiation and fusion by targeting the 3′ untranslated rgion (UTR) of Igf2bp3. Notably, the impacts of miR-6240 mimics on myoblast proliferation, differentiation, and fusion can be effectively counteracted by the overexpression of Igf2bp3. Moreover, our findings elucidate a direct interaction between Igf2bp3 and the myoblast fusion factor myomaker (Mymk). Igf2bp3 binds to Mymk to enhance its mRNA stability. This interaction results in increased expression of Mymk and heightened myoblast fusion. Conclusions Our study unveils Igf2bp3 as a novel post-transcriptional regulator of myoblast fusion through the miR-6240/Mymk axis, significantly contributing to our understanding of skeletal muscle development. Graphical Abstracthttps://doi.org/10.1186/s11658-024-00650-1Igf2bp3Myobalst fusionMymkmRNA stabilitymiR-6240Myogenesis |
| spellingShingle | Yuxin Huang Wei Wang Xinhao Fan Xiaoqin Liu Weiwei Liu Zishuai Wang Yixing Li Yalan Yang Zhonglin Tang The miR-6240 target gene Igf2bp3 promotes myoblast fusion by enhancing myomaker mRNA stability Cellular & Molecular Biology Letters Igf2bp3 Myobalst fusion Mymk mRNA stability miR-6240 Myogenesis |
| title | The miR-6240 target gene Igf2bp3 promotes myoblast fusion by enhancing myomaker mRNA stability |
| title_full | The miR-6240 target gene Igf2bp3 promotes myoblast fusion by enhancing myomaker mRNA stability |
| title_fullStr | The miR-6240 target gene Igf2bp3 promotes myoblast fusion by enhancing myomaker mRNA stability |
| title_full_unstemmed | The miR-6240 target gene Igf2bp3 promotes myoblast fusion by enhancing myomaker mRNA stability |
| title_short | The miR-6240 target gene Igf2bp3 promotes myoblast fusion by enhancing myomaker mRNA stability |
| title_sort | mir 6240 target gene igf2bp3 promotes myoblast fusion by enhancing myomaker mrna stability |
| topic | Igf2bp3 Myobalst fusion Mymk mRNA stability miR-6240 Myogenesis |
| url | https://doi.org/10.1186/s11658-024-00650-1 |
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