Analysis of Casein Derived Circular RNA Expression Profile During Response to Heat Stress
QI Ying, XIE Chun-Yan, XING Jing, LI Qiu-Ling*
College of Life Sciences, Langfang Normal University/Hebei Key Laboratory of Animal Diversity/Langfang Key Laboratory of Cell Engineering and Application, Langfang 065000, China
Abstract Heat stress leads to casein reduction in milk which adversely affects milk quality. The study was aimed to identify the casein derived circular RNA (circRNA) involved in heat stress response and to provide new insight for further elucidation of the molecular mechanism of heat stress. In the previous study of this research group, we obtained the genome wide circRNA expression data of mammary gland of cows (Bos taurus) by RNA-seq. Four circRNAs were randomly selected and the back splice junction sites were confirmed by Sanger sequencing. The relative expression level of circRNAs were verified by quantitative real-time PCR, and the results were consistent with that of RNA sequencing. The casein coding genes include casein alpha s1 (CSN1S1), CSN1S2, casein beta (CSN2) and casein kappa (CSN3). The host genes of circRNAs were analyzed and casein derived circRNAs were identified. Moreover, the expression patterns of these circRNAs in different samples and groups were shown by clustering heat map and Venn diagram. Finally, the relative expression level of casein derived circRNAs were analyzed according to the transcripts per million (TPM) data obtained from RNA sequencing. 52 casein derived circRNAs were identified, of which 29, 16 and 7 were derived from CSN1S1, CSN1S2 and CSN2, respectively. No circRNA derived from CSN3 was detected. Compared with the non heat stress group, 5 circRNAs were significantly up-regulated, 3 of which were only expressed in heat stress condition. The casein derived circRNAs identified in this study were significantly differentially expressed in heat stress group compared to the non heat stressed group, which may be involved in the regulation of milk protein synthesis in heat stress response. The study identified the differentially expressed casein derived circRNAs in heat stressed cows, which may contribute to the understanding of the mechanism of casein reduction casused by heat stress.
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