鸭脂肪细胞中circ-FBLN2鉴定与功能预测

Validation and function prediction of circ-FBLN2 in duck adipocytes

Circular RNA(circRNA)是一类在转录过程中形成的首尾相接的环状非编码RNA,其在多种生物学过程以及疾病发生过程中发挥重要作用。基于前期鸭胚原代脂肪细胞分化过程中circRNA测序结果,本研究选择由FBLN2编码的circ-FBLN2进行验证分析。通过PCR扩增和焦磷酸测序技术获得接合序列,采用Rnase R消化实验进一步验证了circ-FBLN2的存在,采用核质分离技术得出circ-FBLN2主要存在于细胞质中,且其在脂肪细胞诱导分化后显著下调,在樱桃谷鸭各组织中的表达具有一定的差异,主要在脂肪和腿肌中表达。生物信息学分析结果显示,circ-FBLN2与预测到的靶标基因主要参与MAPK,Toll样受体,以及细胞因子与受体互作通路等信号通路,揭示其可能通过对靶基因的调控作用间接调控鸭脂肪细胞分化过程。

Circular RNA(circRNA)is a kind of circular non-coding RNA characterized by head to tail connection during the transcription process,which plays an important role in many biological processes and diseases.Based on the results of circRNA sequencing during pre-adipocyte differentiation,circ-FBLN2 encoded by FBLN2 was selected for validation analysis.The conjugated sequence was obtained by PCR amplification and pyrosequencing.And circ-FBLN2 was further verified by RNase R digestion experiment.The results of nuclear and cytoplasmic separation experiment showed that circ-FBLN2 mainly existed in cytoplasm,and its expression level of duck adipocyte was significantly decreased after differentiation.There was a certain difference in the expression level of different tissues in Cherry Valley duck,it was mainly expressed in fat and leg muscle.Bioinformatics analysis showed that the predicted target genes and circ-FBLN2 were involved in MAPK,Toll-like receptor and cytokine-cytokine receptor interaction pathways and other signaling pathways.All these announced that circ-FBLN2 may indirectly regulate the differentiation of duck adipocytes by regulating the target genes.