Molecular Characterizations of FAM13A and Its Functional Role in Inhibiting the Differentiation of Goat Intramuscular Adipocytes through RIG-I Receptor Signaling Pathway
Abstract
:1. Introduction
2. Materials and Methods
2.1. Experimental Animals and Cell Culture
2.2. CDS and 3′UTR Amplification of FAM13A, and Transfection
2.3. RNA Extraction and Real-Time Quantitative Polymerase Chain Reaction
2.4. Preparation of RNAseq Samples
2.5. QC, GO, KEGG Assay Analysis and qRT-PCR Validation
2.6. MTT Measurement
2.7. Oil Red O Staining
2.8. Luciferase Reporter Assay
2.9. Statistical Analysis
3. Results
3.1. Cloning and Bioinformatics Analysis of the FAM13A Gene in Goats
3.2. Goat FAM13A Inhibits Intramuscular Adipocyte Differentiation
3.3. Overexpression of FAM13A Affects mRNA Transcriptional Profiles in Goat Adipocytes
3.4. Overexpression of FAM13A Inhibits Goat Intramuscular Adipocyte Differentiation via the RIG-I-like Receptor Signaling Pathway
3.5. miR-21-5p Negatively Regulates the Expression of FAM13A
3.6. miR-21-5p Promotes the Differentiation of Goat Intramuscular Adipocytes
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Name | Sequences (5′-3′) |
---|---|
U6 | S: TGGAACGCTTCACGAATTTGCG A: GGAACGATACAGAGAAGATTAGC |
C/EBPα | S: CCGTGGACAAGAACAGCAAC A: AGGCGGTCATTGTCACTGGT |
C/EBPβ | S: CAAGAAGACGGTGGACAAGC A: AACAAGTTCCGCAGGGTG |
PPARγ | S: AAGCGTCAGGGTTCCACTATG A: GAACCTGATGGCGTTATGAGAC |
AP2 | S: TGAAGTCACTCCAGATGACAGG A: TGACACATTCCAGCACCAGC |
LPL | S: TCCTGGAGTGACGGAATCTGT A: GACAGCCAGTCCACCACGAT |
SREBP1 | S: AAGTGGTGGGCCTCTCTGA A: GCAGGGGTTTCTCGGACT |
FAM13A | S: AACTGATGTCGGATGGAAGCAA A: CACGAGGAGAAGGCAGGGATAG |
UXT | S: GCAAGTGGATTTGGGCTGTAAC A: ATGGAGTCCTTGGTGAGGTTGT |
ISG15 | S: ATCAATGTGCCTGCTTTCCA A: GGGCTTCCCTTCAAAAGACAG |
RIG-I | S: GAAGAACTTGGGACCGTAACTC A: CACGCTTTCTGAACTGCGATAA |
MDA5 | S:GGAGATGACAGTGATGAGAGTGATG A: ATTATTCCTCGTGCTGACCCTTC |
CXCL10 | S: TCTGCCTTATCCTTCTGACTCTGA A: TTATGCCTCTTTCCGTGTTCG |
TRIM25 | S: GCCTGTGAAGAAGGTTGTGAAAG A: ACCTTGGCGTTGAGAGATGC |
LGP2 | S: AGAGGCATTTAGAGACCGTGG A: CGCTCAGGGTTGTGATGGT |
IRF7 | S: TGACACGCCCATCTTTGACT A: GCCCAGGTAGATGGTGTAGTG |
FAM13A-3′UTR | S: TCAAGGTTTACGCCAGACTA A: TGGTTGCCAAGAGAAGGTGT |
FAM13A | S: GAAGAAACAGTTGTGAGGGTC A: ATCCTCTGTAGATGAGTGCGT |
NC | 5′ UUCUCCGAACGUGUCACGUTT 3′ 5′ ACGUGACACGUUCGGAGAATT 3′ |
Inhibitor NC | 5′ CAGUACUUUUGUGUAGUACAA 3′ |
miR-21-5p mimic | 5′ UAGCUUAUCAGACUGAUGUUGAC 3′ 5′ CAACAUCAGUCUGAUAAGCUAUU 3′ |
miR-21-5p inhibitor | 5′ GUCAACAUCAGUCUGAUAAGCUA 3′ |
si-NC | 5′ UUCUCCGAACGUGUCACGUTT 3′ 5′ ACGUGACACGUUCGGAGAATT 3′ |
FAM13A siRNA | 5′ CCGAGUAUAAGCACAUUAATT 3′ 5′ UUAAUGUGCUUAUACUCGGTT 3′ |
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Li, X.; Ran, L.; Li, Y.; Wang, Y.; Xiong, Y.; Wang, Y.; Xing, J.; Lin, Y. Molecular Characterizations of FAM13A and Its Functional Role in Inhibiting the Differentiation of Goat Intramuscular Adipocytes through RIG-I Receptor Signaling Pathway. Genes 2024, 15, 1143. https://doi.org/10.3390/genes15091143
Li X, Ran L, Li Y, Wang Y, Xiong Y, Wang Y, Xing J, Lin Y. Molecular Characterizations of FAM13A and Its Functional Role in Inhibiting the Differentiation of Goat Intramuscular Adipocytes through RIG-I Receptor Signaling Pathway. Genes. 2024; 15(9):1143. https://doi.org/10.3390/genes15091143
Chicago/Turabian StyleLi, Xuening, Li Ran, Yanyan Li, Yong Wang, Yan Xiong, Youli Wang, Jiani Xing, and Yaqiu Lin. 2024. "Molecular Characterizations of FAM13A and Its Functional Role in Inhibiting the Differentiation of Goat Intramuscular Adipocytes through RIG-I Receptor Signaling Pathway" Genes 15, no. 9: 1143. https://doi.org/10.3390/genes15091143