Selection of reference genes for RT-qPCR analysis in developing chicken embryonic ovary

Wang, Yi; Zhang, Yuqing; Wu, Zi‐Wei; Fang, Ting; Wang, Fang; Zhao, Han; Du, Zhiqiang; Yang, Caixia

doi:10.1007/s11033-023-08280-0

Cited by 2 publications

(1 citation statement)

References 46 publications

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“…Subsequently, the RefFinder software were utilized to comprehensively arrange the outcomes obtained from the aforementioned 4 algorithms, with the aim of identifying the optimal internal reference gene. This method has been widely employed in poultry research to screen and identify numerous highly suitable reference genes ( Mitra et al, 2016 ; Dunislawska et al, 2020 ; Rodríguez‐Hernández et al, 2021 ; Mogilicherla et al, 2022 ; Chen et al, 2023 ; Wang et al, 2023 ). While traditional methods for screening reference genes are commonly used, it is clear that they have limitations in identifying the most optimal candidates from a limited pool of HKGs and a narrow screening range.…”

Section: Introductionmentioning

confidence: 99%

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

Shu,

Hua,

Zheng

et al. 2024

Poultry Science

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Section: Introductionmentioning

confidence: 99%

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

Shu,

Hua,

Zheng

et al. 2024

Poultry Science

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Screening of reliable reference genes for the normalization of RT-qPCR in chicken liver tissues and LMH cells

Chen,

Hua,

Shu

et al. 2024

Sci Rep

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The liver plays a vital role in lipid synthesis and metabolism in poultry. To study the functional genes more effectively, it is essential to screen of reliable reference genes in the chicken liver, including females, males, embryos, as well as the Leghorn Male Hepatoma (LMH) cell line. Traditional reference gene screening involves selecting commonly used housekeeping genes (HKGs) for RT-qPCR experiments and using different algorithms to identify the most stable ones. However, this approach is limited in selecting the best reference gene from a small pool of HKGs. High-throughput sequencing technology may offer a solution to this limitation. This study aimed to identify the most consistently expressed genes by utilizing multiple published RNA-seq data of chicken liver and LMH cells. Subsequently, the stability of the newly identified reference genes was assessed in comparison to previously validated stable poultry liver expressed reference genes and the commonly employed HKGs using RT-qPCR. The findings indicated that there is a higher degree of similarity in stable expression genes between female and male liver (such as LSM14A and CDC40 ). In embryonic liver, the optimal new reference genes were SUDS3 , TRIM33 , and ERAL1 . For LMH cells, the optimal new reference genes were ALDH9A1 , UGGT1 , and C21H1orf174 . However, it is noteworthy that most HKGs did not exhibit stable expression across multiple samples, indicating potential instability under diverse conditions. Furthermore, RT-qPCR experiments proved that the stable expression genes identified from RNA-seq data outperformed commonly used HKGs and certain validated reference genes specific to poultry liver. Over all, this study successfully identified new stable reference genes in chicken liver and LMH cells using RNA-seq data, offering researchers a wider range of reference gene options for RT-qPCR in diverse situations.

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Selection of reference genes for RT-qPCR analysis in developing chicken embryonic ovary

Cited by 2 publications

References 46 publications

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

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

Screening of reliable reference genes for the normalization of RT-qPCR in chicken liver tissues and LMH cells

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