Identification of Targets and Pathways Controlled by the Chicken MicroRNAs miR-10a and miR-143

spleen;chicken;microRNA;luciferase assay;microarray

Hsia, Hung-Ching ; Dr. H. -C. Sunny Liu, Committee Chair,Dr. Bob Petters, Committee Member,Dr. Chad Stahl, Committee Member,Hsia, Hung-Ching ; Dr. H. -C. Sunny Liu ; Committee Chair ; Dr. Bob Petters ; Committee Member ; Dr. Chad Stahl ; Committee Member

University:North Carolina State University

关键词: spleen;    chicken;    microRNA;    luciferase assay;    microarray;   

Others  :  https://repository.lib.ncsu.edu/bitstream/handle/1840.16/942/etd.pdf?sequence=1&isAllowed=y

美国|英语

来源: null

【 摘 要 】

MicroRNAs are small non-coding RNAs that regulate gene expression at the posttranscriptionallevel. The importance of microRNAs in development, tumorogenesis, immunesystem function, and infectious diseases has gradually come to light. MicroRNAs are oftenexpressed in a temporal and spatial manner and regulate specific gene sets to achievephenotypic change. To understand the role of microRNAs in the chicken embryonic spleen,we have profiled microRNA expression at E15 and E20 chicken embryos. The objective ofthe current project is to identify the targets and pathways controlled by two microRNAs,miR-10a and miR-143. Target prediction was carried out by the algorithm miRanda. Eightpredicted targets of each microRNA were subjected to validation using a reporter assay thatincorporates synthetic or retroviral transduced microRNA. Validated targets for miR-143known to have profound functions in apoptosis, T and B cell development and maturation,cancer, or adipocyte differentiation were identified with this approach; for miR-10a,validated targets involved in hematopoiesis, immune cells homing and migration andmegakaryocyte differentiation were identified. Microarray analysis revealed that severaltargets not predicted by the algorithm were upregulated upon treatment with a specificmicroRNA inhibitor. This result suggests that both microRNAs may regulate genes thatcontribute to cancer formation. In addition, miR-10a may be involved in regulating thecomplement system, which is a crucial component of innate immunity; miR-143 may beinvolved in the PPAR/RXR pathway, which regulates glucose and lipid metabolism in cells.Taken together, our results suggest targets and pathways possibly regulated by miR-10a ormiR-143. The inferred functions of the targets and pathways identified in our study aremostly consistent with those from previous studies, thereby providing insights and directionsfor future research into the mechanisms by which miR-10a and miR-143 exert their functionin the developing chick embryo. Furthermore, the RCAS system used in this project alsoproved the usefulness of engineered retrovirus for transducing microRNA. Furtherdevelopment of this tool may aid microRNA research and further contribute to understandingof the role of microRNAs in vertebrate development.