Abstract: 【Objective】The ZBED6 gene is a transcription factor that regulates muscle growth and development. In order to investigate the regulatory mechanism of ZBED6 in myocardium growth and development, this study was designed to compare the transcriptome of cardiac tissue obtained from Bama miniature pig between the ZBED6 knockout group (ZBED6-KO) and the wild type group (ZBED6-WT) and to find out the effect of ZBED6 gene knockout on the development and gene expression profile of pig heart tissue. 【Method】 The phenotypic characteristics of ZBED6-KO and ZBED6-WT pig tissues were analyzed by t-test and the expression level of the target gene IGF2 of the ZBED6 gene was quantified by real-time quantitative PCR (q-PCR). Comparing the differences in histological level of tissue structure were performed by paraffin section method. The total RNA was isolated from ZBED6-KO group and ZBED6-WT group. RNA-Seq analysis was performed on Illumina Hiseq 2000 platform. The differentially expressed genes between ZBED6-KO and ZBED6-WT heart tissues were screened by the method of RNA-Seq Pig Sus_scrofa10.2 was used as the reference sequence. Differentially expressed genes were enriched and analyzed with IPA software. 9 differentially expressed genes were randomly selected to verify the reproducibility of RNA-Seq results by q-PCR. 【Result】The heart of ZBED6-KO pig had a larger measured data in weight than ZBED6-WT, which also had a higher expression profile of IGF2 than the latter. Compared with ZBED6-WT pig, ZBED6-KO pig had wide muscle fiber width and less connective tissue. The results suggested that the knockout of ZBED6 gene could promote the growth and development of porcine heart. The sequencing results showed that at least 10 G data were obtained and the clean ratio, Q30 data were more than 90%, of which 62.8% -80.1% of the reads mapped to the pig genome. All of these indicated sequencing was well saturation and the sequencing data was reliable. By analyzing sequencing data, 184 different genes were indicated which contained 114 up-regulated genes and 70 down-regulated genes. Among the 184 genes, there were 141 intact function annotation genes and 43 non-annotated genes. Differential gene hierarchical clustering analysis showed that, the similar expression pattern was detected in ZBED6-KO group (x1, x3, x6), which was also happen to ZBED6-WT group (x2, x4, x5). GO and IPA enrichment analysis obtained 13 significant GO terms, 33 pathways. The difference gene was mainly enriched in the immune response, muscle development, RhoA signal and other related channels. The expression pattern of 9 differentially expressed genes was consistent with the results of RNA-Seq analysis. 【Conclusion】In this study, ZBED6-KO Bama miniature pig was first time to use as the model. We used RNA-Seq technique to explore the effects of ZBED6 gene knockout on cardiac development, which enriched the research of ZBED6 on myocardium development and function.

Key words: ZBED6, gene knockout, RNA-Seq, myocardial development, differentially expressed genes