AGRICULTURAL RESEARCH COMMUNICATION CENTRE www.arccjournals.com/www.ijaronline.in *Corresponding author’s e-mail: vetraja86@gmail.com. Indian J. Anim. Res., 51 (5) 2017 : 807-812 Print ISSN:0367-6722 / Online ISSN:0976-0555 Evaluation of function of RICTOR gene in gga-miR-142-3p knockdowned developing chicken embryo Raja Paramasivam*, Prabhakar Tembhurne, Maneshkumar Pannerselvam, Vijay Ingle, Nitin Kurkure and Dewananad Kalorey Department of Veterinary Microbiology and Animal Biotechnology, Teaching and Research Cell, Nagpur Veterinary College, Nagpur-440 001, Maharashtra, India. Received: 03-02-2016 Accepted: 04-05-2016 DOI:10.18805/ijar.10983 ABSTRACT MicroRNAs (miRNAs) are a large classes of endogenous non coding RNAs of about 18-21 nucleotides long. It regulates the gene expression post-transcriptionally by binding partial or complete complementary sites in 3’ UTRs. The miR-142- 3p was highly expressed in all native cell lineages of haematopoietic organs. It has been reported that miR-142-3p was highly expressed in embryonic day 15 and day 20 in spleen and bursa of Fabricius of chicken. The present study was planned to knock down the gga-miR142-3p in chicken embryonic developmental stages for identification of its role in morphogenesis at embryonic spleen and bursa of Fabricius development. Knock down of miR-142-3p in the 14 days old embryonic eggs was carried out by intravenous inoculation of LNA modified miR-142-3p inhibitor. Organs were harvested from the embryos at the embryonic day of 20 and analyzed for gross pathological, histopathological changes and RICTOR gene expressions have been analyzed using SYBR Green qPCR technology. The results indicated that the RICTOR gene was upregulated in all the immune organs and visceral organs except kidney. The over expression of RICTOR gene lead to defective actin reorganization in the cells that will affect the change in the cell structural integrity and cell survival. Over expression of RICTOR gene is associated with increased proliferation of cells along with invasion of tumor cells. Key words: Chicken, Knock down of ggi-miR-142-3p, RICTOR, Structural integrity. Abbreviations: PBS: Phosphate Buffer Saline; H&E: Haematoxylin and Eosin; qPCR: Quantitative Real time Polymerase Chain Reaction. INTRODUCTION Vertebrates have more than 30,000 genes but only few genes are involved in governing of cell behavior and functions. Gene functions cannot be studied without the knowledge of developmental biology but until now only some of the gene functions were analyzed experimentally. The biological functional aspect of gene regulation is controlled by various non coding RNAs. Among them, four types of silencing-related small RNAs have been found in animals over the past few years: small interfering RNAs (siRNAs), microRNAs (miRNAs), repeat associated small interfering RNAs (rasiRNAs) and Piwi-interacting RNAs (piRNAs). Among these, micro RNA plays a major role in the regulation of gene expression and it controls up to 30% of the genome (Bartel, 2004; Sandhu et al., 2011). Some of the miRNAs (miR-125b, miR-142-3p and miR-21) are highly expressed during developmental stages of embryo and in immune organs viz., bursa of Fabricius and spleen during day 15 and 20 which involve a variety of cell proliferation, differentiation and migration of T and B cells. Micro RNA profiling in chicken embryo at day 20 revealed higher expression of gga-miR-142-3p in immune organs such as spleen (5.9%) and bursa (9.7%) as compared to 15 day old embryos (Hicks et al., 2008 and 2009). Micro RNAs interact with mRNAs and either block protein translation or lead to protein degradation (Bartel, 2004). Micro RNAs are typically not important in the early stages of development, but they are extremely important during the later stages of embryonic development, when they regulate cell differentiation and proliferation (Wienholds et al., 2005). RICTOR is the one of the components of the mTOR2 complex. The function of the mTOR2 is mainly actin cytoskeleton reorganization and RICTOR is required for the stability and activity of the mTOR2. RICTOR is the key component of the phosphotidylinositol-3-kinase (P13K)/ mTOR/AKT signaling pathways while regulating the beta cell proliferation and cell size. Activation of P13K or AKT signaling pathways is found to be associated with growth and proliferation of cells and cell survival (Gu et al., 2011). The present study was planned to identify the role of gga-miR-142-3p in the embryonic organs via knocking down of the gga-miR-142-3p in the embryonic developments. The knock down of gga-miR-142-3p was performed by intravenous inoculation of anti-miR-142-3p LNA probe. The