人 NPTX1 相关 H3K27ac 富集的非保守性增强子鉴定.

Objective To explore and identify H3K27ac enriched non-conserved enhancers near human NPTX1, improve the annotation of NPTX1-related regulatory regions, and provide a corresponding theoretical basis for the study of NPTX1-related diseases and mutations in non-coding regions. Methods A total of 39 H3K27ac ChIP-seq data involving 7 tissues were downloaded from the CistromeDB database and analyzed. Peaks in the range of 100 kb on both sides of NPTX1 were selected. Fragments’ sequence conservation among human, mouse and zebrafish was analyzed by ECR browser. The fragments were inserted into a dual luciferase validation vector, and dual luciferase assays were performed in human neural-derived cell line SH-SY5Y and human kidney-derived cell line 293T to detect enhancers’ activity. Results We obtained 5 H3K27ac-enriched fragments. The DNA sequences of each fragment were not conserved among human, mouse and zebrafish. The ratio of dual luciferase activity of NPTX1-E1-P was significantly higher than that of the negative control in SH-SY5Y, but it was about 40% lower in 293T cells. Conclusion Among the H3K27ac-enriched fragments near NPTX1, NPTX1-E1-P has enhancer activity in human neurogenic cell line SH-SY5Y, and low sequence conservation among species. Compared with the human neurogenic cell line SH-SY5Y, its enhancer activity is decreased in human kidney-derived cell line 293T. [ABSTRACT FROM AUTHOR]