selection and validation of reference genes for mirna expression studies during porcine pregnancy选择和验证的参考基因猪怀孕期间microrna的表达研究.pdfVIP

Selection and Validation of Reference Genes for miRNA Expression Studies during Porcine Pregnancy 1 2 1 1,2 Jocelyn M. Wessels , Andrew K. Edwards , Candace Zettler , Chandrakant Tayade * 1 Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, Canada, 2 Department of Biomedical and Molecular Sciences, Queen’s University, Kingston, Canada Abstract MicroRNAs comprise a family of small non-coding RNAs that modulate several developmental and physiological processes including pregnancy. Their ubiquitous presence is confirmed in mammals, worms, flies and plants. Although rapid advances have been made in microRNA research, information on stable reference genes for validation of microRNA expression is still lacking. Real time PCR is a widely used tool to quantify gene transcripts. An appropriate reference gene must be chosen to minimize experimental error in this system. A small difference in miRNA levels between experimental samples can be biologically meaningful as these entities can affect multiple targets in a pathway. This study examined the suitability of six commercially available reference genes (RNU1A, RNU5A, RNU6B, SNORD25, SCARNA17, and SNORA73A) in maternal-fetal tissues from healthy and spontaneously arresting/dying conceptuses from sows were separately analyzed at gestation day 20. Comparisons were also made with non-pregnant endometrial tissues from sows. Spontaneous fetal loss is a prime concern to the commercial pork industry. Our laboratory has previously identified deficits in vasculature development at maternal-fetal interface as one of the major participating causes of fetal loss. Using this well-established model, we