Small non-coding RNA profiling and the role of piRNA pathway genes in the protection of chicken primordial germ cells

BMC Genomics

Research Article

Hojin Jung¹  Seung-Jae Noh¹  Myunghee Jung¹  Young Min Kim²  Jae Yong Han²  Hong Jo Lee²  Sang In Lee²  Yoon Ah Sohn²  Deivendran Rengaraj³  Tae Sub Park⁴ 

[1] Codes Division, Insilicogen, Inc, 441-813, Suwon, Korea;Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, College of Agriculture and Life Sciences, Seoul National University, 151-921, Seoul, Korea;Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, College of Agriculture and Life Sciences, Seoul National University, 151-921, Seoul, Korea;Department of Animal Science and Technology, Chung-Ang University, 456-756, Anseong, Korea;Graduate School of International Agricultural Technology and Institute of Green-Bio Science and Technology, Seoul National University, 232-916, Pyeongchang-gun, Gangwon-do, Korea;

关键词: Aves;    Non-coding RNA;    piRNA;    Primordial Germ Cells;   

DOI  :  10.1186/1471-2164-15-757

 received in 2014-05-16, accepted in 2014-08-29,  发布年份 2014

来源: Springer

【 摘 要 】

BackgroundGenes, RNAs, and proteins play important roles during germline development. However, the functions of non-coding RNAs (ncRNAs) on germline development remain unclear in avian species. Recent high-throughput techniques have identified several classes of ncRNAs, including micro RNAs (miRNAs), small-interfering RNAs (siRNAs), and PIWI-interacting RNAs (piRNAs). These ncRNAs are functionally important in the genome, however, the identification and annotation of ncRNAs in a genome is challenging. The aim of this study was to identify different types of small ncRNAs particularly piRNAs, and the role of piRNA pathway genes in the protection of chicken primordial germ cells (PGCs).ResultsAt first, we performed next-generation sequencing to identify ncRNAs in chicken PGCs, and we performed ab initio predictive analysis to identify putative piRNAs in PGCs. Then, we examined the expression of three repetitive sequence-linked piRNAs and 14 genic-transcript-linked piRNAs along with their linked genes using real-time PCR. All piRNAs and their linked genes were highly expressed in PGCs. Subsequently, we knocked down two known piRNA pathway genes of chicken, PIWI-like protein 1 (CIWI) and 2 (CILI), in PGCs using siRNAs. After knockdown of CIWI and CILI, we examined their effects on the expression of six putative piRNA-linked genes and DNA double-strand breakage in PGCs. The knockdown of CIWI and CILI upregulated chicken repetitive 1 (CR1) element and RAP2B, a member of RAS oncogene family, and increased DNA double-strand breakage in PGCs.ConclusionsOur results increase the understanding of PGC-expressed piRNAs and the role of piRNA pathway genes in the protection of germ cells.

【 授权许可】

Unknown   
© Rengaraj et al.; licensee BioMed Central Ltd. 2014. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.