CircRNA related to subcutaneous fat of pigs and application thereof

A technology of subcutaneous fat and pig skin, applied in DNA/RNA fragments, recombinant DNA technology, determination/inspection of microorganisms, etc., can solve problems such as unclear

Active Publication Date: 2018-11-27
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether circRNA is involved in fat deposition and metabolism is still unclear

Method used

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  • CircRNA related to subcutaneous fat of pigs and application thereof
  • CircRNA related to subcutaneous fat of pigs and application thereof
  • CircRNA related to subcutaneous fat of pigs and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 circRNA sequencing and differentially expressed circRNA selection

[0041] 1.1 Experimental animals:

[0042]In this experiment, large white pigs and Laiwu pigs with obvious differences in fat deposition were used as materials. They were raised in Laiwu Daqian Agriculture and Animal Husbandry Co., Ltd., grown and fattened under the same feeding conditions and environment, and fed according to the nutritional requirements standard (National Research Council, NRC, 1998) To feed the diet, choose 3 large white pigs (about 100kg) and 3 Laiwu pigs (about 35kg) each with a similar weight in the species, which are 180 days old, healthy and in good condition.

[0043] 1.2 Sample collection:

[0044] After slaughtering the experimental pigs, the subcutaneous adipose tissue was quickly separated. To reduce RNA degradation, all processes were performed on ice. Afterwards, the tissue was cut into small pieces with sterile scissors, put into 5mL cryopreservation tubes, f...

Embodiment 2

[0072] Example 2 In-depth analysis of circRNA

[0073] 2.1 Functional analysis of differentially expressed circRNA:

[0074] After obtaining the differentially expressed circRNAs, we used the information of the circRNA source genes to perform GO enrichment analysis on the differentially expressed circRNAs. The number of differential circRNAs included in each GO entry was counted, and the significance of differential circRNA enrichment in each GO entry was calculated using the hypergeometric distribution test method. KEGG is an important public database related to Pathway. Using the KEGG database to perform Pathway analysis on differential circRNA-derived genes, you can find Pathway entries that enrich differential circRNA-derived genes, and look for potential differential circRNAs in different samples that may be related to changes in cellular pathways. .

[0075] 2.2 circRNA-miRNA-mRNA interaction network diagram

[0076] The circRNA_11897 down-regulated in Laiwu subcutane...

Embodiment 3

[0079] Embodiment 3 real-time fluorescent quantitative PCR verification

[0080] Select circRNA_11897, 2 miRNAs (ssc-miR-27a, ssc-miR-27b-3p) and SCD gene in the target gene of circRNA_11897, and set 20 biological replicates for each (Large White pig and Laiwu pig 20 cases each), Glyceraldehyde-3-phosphate dehydrogenase (glyceraldehyde-3-phosphate dehydrogenase, GAPDH) gene and 5S rRNA were used as internal references, respectively, and qRT-PCR method was used to verify the expression level of the genes.

[0081] 3.1 Primer design:

[0082] circRNA_11897 amplification:

[0083] Forward primer: 5'-TAACCCATACCGGTCCTTGC-3' (SEQ ID NO.2)

[0084] Reverse primer: 5'-ATAACAAGCTTGCTGGCCTCT-3' (SEQ ID NO.3)

[0085] SCD amplification:

[0086] Forward primer: 5'-CCTTTATTCTTCTCTAACCCGT-3' (SEQ ID NO.4)

[0087] Reverse primer: 5'-TCAGTGAGCAGAGAGACTTGTG-3' (SEQ ID NO.5)

[0088] ssc-miR-27a amplification:

[0089] Specific primer: 5'-TTCACAGTGGCTAAGTTCCGC-3' (SEQ ID NO.6)

[009...

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Abstract

The invention relates to a circRNA related to subcutaneous fat of pigs and an application thereof, and specifically relates to a circRNA_11897 related to subcutaneous fat of pigs and the application thereof. The invention identifies and analyzes the circRNA of the subcutaneous fat of large white pigs and Laiwu pigs, adopts a systematic bioinformatic method for performing functional analysis on thedifferential expression circRNA and finds the circRNA_11897 and the genes ssc-miR-27a, ssc-miR-27b-3p and SCD related to the circRNA_11897. The experiment proves that the expression quantity of the circRNA_11897 and the genes has a close relation with the subcutaneous fat of pigs. The invention provides a new theoretical basis and a potential target for improving the quality of pork and preventing the happening of metabolic diseases caused by dyslipidemia.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a pig subcutaneous fat-related circular RNA and its application, more specifically to circRNA_11897 and its application. Background technique [0002] The quality of pork is closely related to human health. Excessive deposition of subcutaneous fat will increase the content of cholesterol and reduce the quality of pork. Consumers will easily lead to obesity after ingestion. Therefore, understanding the molecular mechanism of fat deposition will help improve the quality of pork. Pork quality and human health. Large white pig, as a typical lean pig, has low subcutaneous fat; while Laiwu pork has bright red color, good water holding capacity, high fat content, and excellent meat quality. Therefore, the significant difference in fat deposition between Large White and Laiwu pigs provides a good model for the study of fat metabolism. [0003] circRNA is a kind of circular endogeno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/113
CPCC12N15/113C12N2310/10C12Q1/6888C12Q2600/124C12Q2600/158C12Q2600/178C12N2310/532
Inventor 苗向阳李嫒解领丽黄万龙张秀秀
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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