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.

ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
    Porcine embryo collection and in vitro maturation
    Parthenogenetic activation and embryo developmentin vitro
    Double stranded RNA (dsRNA) injection and embryocollection
    Immunocytochemistry (ICC)
    Real time reverse-transcription polymerase chainreaction (real time RT-PCR)
    Quantification of m6A RNA methylation
    Statistical analysis
RESULTS
    Characterization of hnRNPA2/B1 during porcineparthenotes
    Effect of hnRNPA2/B1 knock-down during porcineearly embryo development
    hnRNPA2/B1 knock-down alters global geneexpression
DISCUSSION
REFERENCES

• Jeongwoo Kwon(Primate Resources Center (PRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56216, Korea)
• Yu-Jin Jo(Primate Resources Center (PRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56216, Korea)
• Seung-Bin Yoon(Primate Resources Center (PRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56216, Korea)
• Hyeong-ju You(Primate Resources Center (PRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56216, Korea)
• Changsic Youn(Primate Resources Center (PRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56216, Korea)
• Yejin Kim(Primate Resources Center (PRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56216, Korea)
• Jiin Lee(Primate Resources Center (PRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56216, Korea)
• Nam-Hyung Kim(Guangdong Provincial Key Laboratory of Large Animal models for Biomedicine, Wuyi university, Jiangmen 529020, China) Corresponding Author
• Ji-Su Kim(Primate Resources Center (PRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56216, Korea) Corresponding Author