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| 1 | miR-25 modulates triacylglycerol and lipid accumulation in goat mammary epithelial cells by repressing PGC-1beta显示文摘Background: The goat(Caprahircus) is one of the most important livestock animals. Goat milk fat is an important component in the nutritional quality of goat milk. Growing evidence points to the critical roles of microRNAs(miRNAs) in lipid metabolism.Results: Using a highly sensitive method of S-poly(T) plus for miRNAs detection, we analyze the expression patterns of 715 miRNAs in goat mammary gland tissues at different stages of lactation. We observed that miR-25 expression had an inverse relationship with milk production. Overexpression of miR-25 significantly repressed triacylglycerol synthesis and lipid droplet accumulation. To explore the regulatory mechanism of miR-25 in milk lipid metabolism,we analyzed its putative target genes with bioinformatics analysis followed by 3′-UTR assays. Peroxisome proliferative activated receptor gamma coactivator 1 beta(PGC-1 beta), a key regulator of lipogenics was identified as a direct target of miR-25 with three specific sites within its 3′-UTR. In addition, miR-25 mimics in goat mammary epithelial cells reduced the expressions of genes involved in lipid metabolism.Conclusions: Taken together, our results show miR-25 is potentially involved in lipid metabolism and we reveal the function of the miR-25/PGC-1 beta regulatory axis during lactation. | Liuan Ma Huiling Qiu Zhi Chen Li Li Yan Zeng Jun Luo Deming Gou | 2018 | Journal of Animal Science and Biotechnology2018，9，4： | 9 |
| 2 | Identification of circulating miRNA biomarkers based on global quantitative real-time PCR profiling显示文摘MicroRNAs (miRNAs) are small noncoding RNAs (18-25 nucleotides) that regulate gene expression at the post-transcriptional level. Recent studies have demonstrated the presence of miRNAs in the blood circulation. Deregulation of miRNAs in serum or plasma has been associated with many diseases including cancers and cardiovascular diseases, suggesting the possible use of miRNAs as diagnostic biomarkers. However, the detection of the small amount of miRNAs found in serum or plasma requires a method with high sensitivity and accuracy. Therefore, the current study describes polymerase chain reaction (PCR)-based methods for measuring circulating miRNAs. Briefly, the procedure involves four major steps: (1) sample collection and preparation; (2) global miRNAs profiling using quantitative real-time PCR (qRT-PCR); (3) data normalization and analysis; and (4) selection and validation of miRNA biomarkers. In conclusion, qRT-PCR is a promising method for profiling of circulating miRNAs as biomarkers. | Kang Kang Xiao Peng Jun Luo Deming Gou | 2012 | Journal of Animal Science and Biotechnology2012，3，2： | 3 |
| 3 | A direct real-time polymerase chain reaction assay for rapid high-throughput detection of highly pathogenic North American porcine reproductive and respiratory syndrome virus in China without RNA purification显示文摘Background:Porcine reproductive and respiratory syndrome virus(PRRSV),and particularly its highly pathogenic genotype(HP-PRRSV),have caused massive economic losses to the global swine industry.Results:To rapidly identify HP-PRRSV,we developed a direct real-time reverse transcription polymerase chain reaction method(dRT-PCR) that could detect the virus from serum specimen without the need of RNA purification.Our dRT-PCR assay can be completed in 1.5 h from when a sample is received to obtaining a result.Additionally,the sensitivity of dRT-PCR matched that of conventional reverse transcription PCR(cRT-PCR) that used purified RNA.The lowest detection limit of HP-PRRSV was 6.3 TCID_(50) using dRT-PCR.We applied dRT-PCR assay to 144 field samples and the results showed strong consistency with those obtained by cRT-PCR.Moreover,the dRT-PCR method was able to tolerate 5-20%(v/v) serum.Conclusions:Our dRT-PCR assay allows for easier,faster,more cost-effective and higher throughput detection of HP-PRRSV compared with cRT-PCR methods.To the best of our knowledge,this is the first report to describe a real-time RT-PCR assay capable of detecting PRRSV in crude serum samples without the requirement for purifying RNA.We believe our approach has a great potential for application to other RNA viruses. | Kang Kang Keli Yang Jiasheng Zhong Yongxiang Tian Limin Zhang Jianxin Zhai Li Zhang Changxu Song Christine Yuan Gou Jun Luo Deming Gou | 2015 | Journal of Animal Science and Biotechnology2015，6，1： | 2 |
| 4 | An improved Tet-on system in micro RNA overexpression and CRISPR/Cas9-mediated gene editing显示文摘Background: Tetracycline(Tet)-regulated expression system has become a widely applied tool to control gene activity. This study aimed to improve the Tet-on system with superior regulatory characteristics.Results: By comprehensively comparing factors of transactivators, Tet-responsive elements(TREs), orientations of induced expression cassette, and promoters controlling the transactivator, we developed an optimal Tet-on system with enhanced inducible efficiency and lower leakiness. With the system, we successfully performed effective inducible and reversible expression of micro RNA, and presented a more precise and easily reproducible fine-tuning for confirming the target of a mi RNA. Finally, the system was applied in CRISPR/Cas9-mediated knockout of nuclear factor of activated T cells-5(NFAT5), a protective transcription factor in cellular osmoregulation.Conclusions: This study established an improved Tet-on system for powerful and stringent gene regulation in functional genetic studies. | Kang Kang Lian Huang Qing Li Xiaoyun Liao Quanjin Dang Yi Yang Jun Luo Yan Zeng Li Li Deming Gou | 2019 | Journal of Animal Science and Biotechnology2019，10，4： | 2 |
| 5 | Knockdown of flotillin-2 inhibits lung surfactant secretion by alveolar type Ⅱ cells显示文摘 | Narendranath Reddy Chintagari Deming Gou Lin Liu | 2008 | Cell Research2008，18，6： | 1 |
| 6 | Gene silencing in mammalian cells by PCR based short hairpin RNA显示文摘 | Nili Jin Lin Liu | 2003 | FEBS letters2003，548，： | 1 |
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