Chen Cai, Zheng Yao, Wang Mengli, Murani Eduard, D'Alessandro Enrico, Moawad Ali Shoaib, Wang Xiaoyan, Wimmers Klaus, Song Chengyi
College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.
Leibniz Institute for Farm Animal Biology (FBN), 18196 Dummerstorf, Germany.
Animals (Basel). 2021 Jun 23;11(7):1871. doi: 10.3390/ani11071871.
The genetic diversity of the axis genes and their association with the variation of gene expression and phenotypic traits, principally represented by SNPs, have been extensively reported. Nevertheless, the impact of retrotransposon insertion polymorphisms (RIPs) on the axis gene activity has not been reported. In the present study, bioinformatic prediction and PCR verification were performed to screen RIPs in four axis genes (, , and ). In total, five RIPs, including one SINE RIP in intron 3 of , one L1 RIP in intron 7 of , and three SINE RIPs in intron 1, intron 5 and intron 9 of , were confirmed by PCR, displaying polymorphisms in diverse breeds. Dual luciferase reporter assay revealed that the SINE insertion in intron 1 of significantly repressed the promoter activity in PK15, Hela, C2C12 and 3T3-L1 cells. Furthermore, qPCR results confirmed that this SINE insertion was associated with a decreased expression of in the leg muscle and longissimus dorsi, indicating that it may act as a repressor involved in the regulation of expression. In summary, our data revealed that RIPs contribute to the genetic variation of axis genes, whereby one SINE RIP in the intron 1 of may decrease the expression of by acting as a repressor.
轴基因的遗传多样性及其与基因表达变异和表型性状（主要由单核苷酸多态性代表）的关联已被广泛报道。然而，逆转录转座子插入多态性（RIPs）对轴基因活性的影响尚未见报道。在本研究中，进行了生物信息学预测和PCR验证，以筛选四个轴基因（、、和）中的RIPs。通过PCR共确认了五个RIPs，包括在内含子3中的一个SINE RIP、在内含子7中的一个L1 RIP以及在内含子1、内含子5和内含子9中的三个SINE RIPs，在不同品种中表现出多态性。双荧光素酶报告基因检测显示，内含子1中的SINE插入显著抑制了PK15、Hela、C2C12和3T3-L1细胞中的启动子活性。此外，qPCR结果证实，这种SINE插入与腿部肌肉和背最长肌中表达的降低有关，表明它可能作为一种阻遏物参与表达的调控。总之，我们的数据表明RIPs促成了轴基因的遗传变异，其中内含子1中的一个SINE RIP可能通过作为阻遏物降低的表达。
