Validation and function prediction of circ-FBLN2 in duck adipocytes

doi:10.3969/j.issn.1004-1524.2021.04.05

Acta Agriculturae Zhejiangensis ›› 2021, Vol. 33 ›› Issue (4): 602-609.DOI: 10.3969/j.issn.1004-1524.2021.04.05

• Animal Science • Previous Articles Next Articles

Validation and function prediction of circ-FBLN2 in duck adipocytes

HU Xiaodan(), WANG Laidi, YANG Ting, BAI Hao, JIANG Yong, WANG Zhixiu, WANG Shasha, CHEN Guohong, CHANG guobin^*()

College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China

Received:2020-07-17 Online:2021-04-25 Published:2021-04-25
Contact: CHANG guobin

Abstract

Abstract:

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.

Key words: circ-FBLN2, Cherry Valley duck, adipocyte, tissues expression

CLC Number:

S834

HU Xiaodan, WANG Laidi, YANG Ting, BAI Hao, JIANG Yong, WANG Zhixiu, WANG Shasha, CHEN Guohong, CHANG guobin. Validation and function prediction of circ-FBLN2 in duck adipocytes[J]. Acta Agriculturae Zhejiangensis, 2021, 33(4): 602-609.

Figures/Tables 10

Table 1 Design of experimental primers

引物名称Primer names	序列sequences (5'-3')	长度Product length/bp
circ-FBLN2 divergent primer	TTCAGTGCCACAACCTCTCC	194
	TCAGCTCCGAATCTGCAGTG
circ-FBLN2 convergent primer	GGAGAGGTTGTGGCACTGAA	1314
	CACTGCAGATTCGGAGCTGA
FBLN2	TCCACCATCAAGGAGAACAG	217
	CACAGCAGGACAGCATCACA
GAPDH	CACAGCCACACACGAAGACA
	CCTTAGCCAGCCCCAGTAGA	106

Fig.1 circ-FBLN2 agarose gel electrophoresis

Fig.2 circ-FBLN2 Pattern maps and conjugated sequence sequencing peaks

Fig.3 The relative expression of circ-FBLN2 in the treatment of RNase R and control group

Fig.4 circ-FBLN2 subcellular localization

Fig.5 circ-FBLN2 and major gene expression during adipocyte differentiation in Cherry Valley duck Different letter of the same gene means siginificant difference(P<0.05).

Fig.6 Expression of circ-FBLN2 gene in different tissues of Cherry Valley duck

Fig.7 ceRNA(circ-FBLN2-miRNA-mRNA) network diagram

Fig.8 GO analysis of 107 target genes

Fig.9 KEGG analysis of 107 target genes

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Validation and function prediction of circ-FBLN2 in duck adipocytes

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