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CircRNA_14707 and application thereof in molecular assistant breeding

A technology of reagents and technologies, applied in circRNA_14707 and its application in molecular-assisted breeding, 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_14707 and application thereof in molecular assistant breeding
  • CircRNA_14707 and application thereof in molecular assistant breeding
  • CircRNA_14707 and application thereof in molecular assistant breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 circRNA sequencing and differentially expressed circRNA selection

[0042] 1.1 Experimental animals:

[0043] 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.

[0044] 1.2 Sample collection:

[0045]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

[0069] Example 2 In-depth analysis of circRNA

[0070] 2.1 Functional analysis of differentially expressed circRNA:

[0071] 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. Using the KEGG database to perform Pathway analysis of differential circRNA source genes, Pathway entries enriched for differential circRNA source genes can be found, and potential differential circRNAs in different samples may be related to changes in cellular pathways.

[0072] 2.2 circRNA-miRNA-mRNA interaction network diagram

Embodiment 3

[0074] Embodiment 3 real-time fluorescent quantitative PCR verification

[0075] Select circRNA_14707, 2 miRNAs (ssc-miR-874, ssc-miR-1285), SCD gene, and SMAD7 gene in the target gene of circRNA_14707, and each set up 20 biological replicates (20 cases for Large White pig and 20 cases for Laiwu pig) , glyceraldehyde-3-phosphate dehydrogenase (glyceraldehyde-3-phosphate dehydrogenase, GAPDH) gene and 5S rRNA were used as internal references, and the qRT-PCR method was used to verify the expression level of the gene.

[0076] 3.1 Primer design:

[0077] circRNA_14707 amplification:

[0078] Forward primer: 5'-TCACCTCCGAAGACTCCAGAA-3'(SEQ ID NO.2)

[0079] Reverse primer: 5'-GCTTCCACCTCGCATTTTGC-3' (SEQ ID NO.3)

[0080] SCD amplification:

[0081]Forward primer: 5'-CCTTTATTCTTCTCTAACCCGT-3' (SEQ ID NO.4)

[0082] Reverse primer: 5'-TCAGTGAGCAGAGAGACTTGTG-3' (SEQ ID NO.5)

[0083] SMAD7 amplification:

[0084] Forward primer: 5'-CAGCTCTACAAGAAGCCGA -3' (SEQ ID NO.6)

[0...

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Abstract

The invention relates to circRNA_14707 and an application thereof in molecular assistant breeding. By means of high-throughput sequencing on expression conditions of circRNA in subcutaneous fat tissues of a big white pig and a Laiwu pig and integrated analysis performed by combining previous miRNA and mRNA of a laboratory, a result shows that differential expressions of the circRNA_14707 and related miRNA are quite obvious and the circRNA_14707 and related miRNA can serve as molecular markers for detecting the subcutaneous fat contents of the pigs. The circRNA_14707 has a very good applicationprospect in the fields of predicating or assisted predicting pork quality and breeding pigs with different muscle qualities and the like.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to circRNA_14707 and its application in molecular assisted breeding. Background technique [0002] circRNA is a kind of circular endogenous molecule produced by special alternative splicing. It is a covalently closed circular structure. It is ubiquitous in animal and plant tissues and cells. It is abundant, stable, conserved and tissue-specific. . Studies have confirmed that circRNAs are involved in the regulation of animal economic traits. Liang et al. identified 149circRNAs in the skeletal muscle tissue of Guizhou minipigs, which are involved in biological processes such as muscle development, muscle contraction, and chromatin modification, revealing that circRNAs are involved in the regulation of muscle development and muscle contraction. However, whether circRNA is involved in fat deposition and metabolism remains unclear. [0003] With the rapid development of biotechnology, usi...

Claims

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Application Information

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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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