Screening of reliable reference genes for the normalization of RT-qPCR in chicken oviduct tract

文献类型： 外文期刊

第一作者： Shu, Xin

作者： Shu, Xin;Zheng, Xiaotong;Chen, Ziwei;Zhang, Jilong;Zhuang, Wuchao;Chen, Jianfei;Shu, Xin;Zheng, Xiaotong;Chen, Ziwei;Chen, Jianfei;Hua, Guoying

作者机构：

关键词： chicken; RT-qPCR; stable reference gene; RNA-seq; oviduct

期刊名称：POULTRY SCIENCE （ 影响因子：3.8； 五年影响因子：4.1 ）

ISSN： 0032-5791

年卷期： 2024 年 103 卷 9 期

页码：

收录情况： SCI

摘要： Utilizing publicly available RNA-seq data to screen for ideal reference genes is more efficient and accurate than traditional methods. Previous studies have identified optimal reference genes in various chicken tissues, but none have specifically focused on the oviduct (including the infundibulum, magnum, isthmus, uterus, and vagina), which is crucial for egg production. Identifying stable reference genes in the oviduct is essential for improving research on gene expression levels. This study investigated genes with consistent expression patterns in the chicken oviduct, encompassing both individual oviduct tract tissues and the entire oviduct, by utilizing multiple RNA-seq datasets. The screening results revealed the discovery of 100 novel reference genes in each segment of oviduct tissues, primarily associated with cell cycle regulation and RNA binding. Moreover, the majority of housekeeping genes ( HKGs ) showed inconsistent expression levels across distinct samples, suggesting their lack of stability under varying conditions. The stability of the newly identified reference genes was assessed in comparison to previously validated stable reference genes in chicken oviduct and commonly utilized HKGs, employing traditional reference gene screening methods. HERPUD2, CSDE1, VPS35, PBRM1, LSM14A , and YWHAB were identified to be suitable novel reference gene for different parts of the oviduct. HERPUD2 and YWHAB were reliable for gene expression normalization throughout the oviduct tract. Furthermore, overexpression and interference assays in DF1 cells showed LSM14A and YWHAB play a crucial role in cell proliferation, highlighting the importance of these newly reference genes for further research. Overall, this study has expanded the options for reference genes in RT-qPCR experiments in different segments of the chicken oviduct and the entire oviduct.

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