원문정보
초록
영어
Heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNPA2/B1) is an N6-methyladenosine (m6A) RNA modification regulator and a key determinant of premRNA processing, mRNA metabolism and transportation in cells. Currently, m6A reader proteins such as hnRNPA2/B1 and YTHDF2 has functional roles in mice embryo. However, the role of hnRNPA2/B1 in porcine embryogenic development are unclear. Here, we investigated the developmental competence and mRNA expression levels in porcine parthenogenetic embryos after hnRNPA2/B1 knock-down. HhnRNPA2/B1 was localized in the nucleus during subsequent embryonic development since zygote stage. After hnRNPA2/B1 knock-down using double stranded RNA injection, blastocyst formation rate decreased than that in the control group. Moreover, hnRNPA2/B1 knock-down embryos show developmental delay after compaction. In blastocyste stage, total cell number was decreased. Interestingly, gene expression patterns revealed that transcription of Pou5f1, Sox2, TRFP2C, Cdx2 and PARD6B decreased without changing the junction protein, ZO1, OCLN, and CDH1. Thus, hnRNPA2/B1 is necessary for porcine early embryo development by regulating gene expression through epigenetic RNA modification.
목차
INTRODUCTION
MATERIALS AND METHODS
Porcine embryo collection and in vitro maturation
Parthenogenetic activation and embryo development in vitro
Double stranded RNA (dsRNA) injection and embryo collection
Immunocytochemistry (ICC)
Real time reverse-transcription polymerase chain reaction (real time RT-PCR)
Quantification of m6A RNA methylation
Statistical analysis
RESULTS
Characterization of hnRNPA2/B1 during porcine parthenotes
hnRNPA2/B1 knock-down alters global gene expression
DISCUSSION
REFERENCES