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Novel circRNA related to intramuscular fat and application thereof

An intramuscular fat and reagent technology, applied in DNA/RNA fragments, recombinant DNA technology, microbial assay/inspection, etc., which can solve problems such as unclearness and complex interaction network.

Pending Publication Date: 2019-04-16
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
Existing studies have shown that one of the main functions of circRNAs is the role of miRNA sponges. However, since most circRNAs have multiple miRNA sponge binding sites, the circRNA-miRNA interaction network is relatively complex, and there are many circRNAs targeting the same miRNA. , or many miRNAs bind to the same circRNA

Method used

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  • Novel circRNA related to intramuscular fat and application thereof
  • Novel circRNA related to intramuscular fat and application thereof
  • Novel circRNA related to intramuscular fat and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 circRNA sequencing and differentially expressed circRNA selection

[0045] 1.1 Experimental animals:

[0046] 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.

[0047] 1.2 Sample collection:

[0048] After slaughtering the experimental pigs, the intramuscular 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,...

Embodiment 2

[0074] Example 2 In-depth analysis of circRNA

[0075] 2.1 Functional analysis of differentially expressed circRNA:

[0076]After the differentially expressed circRNAs were obtained, the GO enrichment analysis was performed on the differentially expressed circRNAs using the information of the circRNA source genes. 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. . There are significant differences in intramuscular fat deposition between Large White pigs and Laiwu pigs. Throug...

Embodiment 3

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

[0080] circRNA_08840, ssc-miR-339-3p in the target gene of circRNA_08840 were selected, and 20 biological replicates were set up for each (20 cases in each of Large White pig and Laiwu pig), and glyceraldehyde-3-phosphate dehydrogenase (glyceraldehyde-3-phosphate dehydrogenase) Phosphate dehydrogenase, GAPDH) gene and 5S rRNA were used as internal references, and qRT-PCR method was used to verify the expression level of the gene.

[0081] 3.1 Primer design:

[0082] circRNA_08840 amplification:

[0083] Forward primer: 5'-cctcggccactgtgttccta-3'(seq id no.2)

[0084] Reverse primer: 5'-cgtggttagtttctgcgttgg-3'(seq id no.3)

[0085] ssc-mir-339-3p:

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

[0087] 3.2 Reverse transcription system:

[0088] Add the following substances into a 0.5ml centrifuge tube: RNA 5μl; Oligo(dT) 2μl; dd H2O (DEPC treated) 4.5μl;

[0089] The above firs...

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Abstract

The invention relates to novel circRNA related to intramuscular fat and application thereof, in particular to circRNA_08840 related to intramuscular fat and new application of an interaction gene ssc-miR-339-3p of circRNA_08840. The two kinds of typical pigs which are large white pigs and Laiwu pigs are selected as research objects, a biological information science method is used for conducting sequencing and data analyzing on circRNA, miRNA and an interaction relation of circRNA and miRNA in the intramuscular fat of the two kinds of pigs, several quite good intramuscular fat content marks arefound, and experimental results show that circRNA_08840 and the interaction gene ssc-miR-339-3p of circRNA_08840 are remarkable in expression difference of intramuscular fat tissue of the two kinds of typical pigs, and novel circRNA is expected to breed pigs with different meat qualities as a mark in molecule auxiliary breeding.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a new circRNA (circRNA_08840) related to intramuscular fat and its application, more specifically to a new application of circRNA_08840 related to intramuscular fat and its interaction gene ssc-miR-339-3p . Background technique [0002] Pig fat is generally divided into subcutaneous fat, intramuscular fat and intermuscular fat. The amount of intermuscular fat is small and similar to subcutaneous fat, so it is generally believed that fat is mainly divided into subcutaneous fat and intramuscular fat. Subcutaneous fat contains high saturated fatty acids, excessive intake will increase serum cholesterol levels, accelerate arteriosclerosis and lead to the occurrence of coronary heart disease and other diseases, so for many years the breeding has been aimed at high lean meat rate and rapid growth, but Accompanied by the decline in pork quality, the main reason is the reduction of ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/113
CPCC12Q1/6888C12Q2600/124C12Q2600/158C12Q2600/178
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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