Novel application of lncRNA and trans-regulation gene WNT11 thereof

A technology of gene and target gene, applied in the field of molecular biology, can solve the problem that the relationship between pig intramuscular fat has not been reported.

Active Publication Date: 2018-05-29
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

WNT family members include WNT1, WNT2, WNT2B, WNT3, WNT3A, WNT4, WNT5A, WNT5B, WNT6, WNT7A, WNT7B, WNT8A, WNT8B, WNT9A, WNT9B, WNT10A, WNT10B, WNT11, WNT16 a total of 19 members, of which WNT11 has Studies have shown that it is related to lung cancer, esophageal cancer, melanoma, and cervical squamous cell carcinoma, but its relationship with pig intramuscular fat has not been reported

Method used

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  • Novel application of lncRNA and trans-regulation gene WNT11 thereof
  • Novel application of lncRNA and trans-regulation gene WNT11 thereof
  • Novel application of lncRNA and trans-regulation gene WNT11 thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Sample collection preparation and experimental design

[0048] After the experimental pigs were slaughtered, the intramuscular fat tissue of the longissimus dorsi was quickly collected, cut into small pieces, filled into a 5mL cryotube, put into liquid nitrogen for freezing, and then transferred to a -80℃ refrigerator for long-term storage for total RNA The experiment set up 3 groups to identify the lncRNA in the intramuscular adipose tissue (D_JN) of the large white pig and the intramuscular adipose tissue (L_JN) of the Laiwu pig, and analyze the gene expression profile of their intramuscular adipose tissue. Each sample is set 3 repetitions.

Embodiment 2

[0049] Example 2 Extraction and quality control of sample total RNA

[0050] Take out the same amount of adipose tissue samples stored at low temperature, and use mirVana according to the instruction manual TM The RNA extraction kit extracts total RNA from each adipose tissue sample, and the separated total RNA samples are stored in a refrigerator at -80°C. Use NanoDrop 2000 spectrophotometer to measure RNA sample concentration and OD260nm / OD280nm value, and control it between 1.9~2.1, use Bioanalyzer 2100 to evaluate the quality of total RNA, and control RIN> = 7 and 28S / 18S> =0.7, use RNase-free DNase I to eliminate potential genomic DNA contamination.

Embodiment 3

[0051] Example 3 cDNA library construction and RNA sequencing

[0052] Strand specific cDNA library

[0053] (1) Ribo-zero kit removes rRNA

[0054] (2) RNA fragmentation

[0055] (3) Synthesis and purification of double-stranded cDNA

[0056] (4) End repair, adding A base

[0057] (5) Sequencing adapter connection

[0058] (6) DNA fragment enrichment and purification

[0059] (7) Library quality inspection

[0060] (8) In this study, a total of 6 cDNA libraries were established, namely D_JN_1, D_JN_2, D_JN_3 (Large white pig intramuscular adipose tissue cDNA library) and L_JN_1, L_JN_2, L_JN_3 (Laiwu pig intramuscular adipose tissue cDNA library).

[0061] RNA-Seq(Illumina Sequence)

[0062] After the library is qualified, the Illumina HiSeqTM 2500 sequencing platform is used to sequence and analyze the cDNA library using Paired-end Sequence. The off-machine data is the raw reads of the original sequencing data.

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Abstract

The invention relates to novel application of lncRNA and a trans-regulation gene WNT11 thereof. The novel application is characterized in that adipose tissue gene expression profiles in large white pigs and Laiwu pigs are compared and analyzed by using a high-throughput sequencing technology, key differential expression lncRNA related with adipogenic differentiation and lipid metabolism is screened out, and expression of signaling pathway family member WNT11 of the lncRNA regulation gene Wnt is displayed by further analysis. According to the invention, comprehensive analysis on mRNA and lncRNAfor regulating procine intramuscular fat deposition is carried out, and certain reference for breeding porcine breeds with high meat quality as well as researching prevention and treatment of lipid metabolism-related diseases is provided.

Description

Technical field [0001] The invention belongs to the technical field of molecular biology, and specifically relates to the new use of down-regulated lncRNA and its trans-regulating gene WNT11, and more specifically to the application of lncRNA and its trans-regulating gene WNT11 in breeding high-quality livestock. Background technique [0002] lncRNA is a type of RNA with a length of more than 200 nt. They do not encode proteins, but participate in the regulation of protein-coding genes at multiple levels (epigenetic regulation, transcription regulation, post-transcriptional regulation, etc.) in the form of RNA, including It plays a key regulatory role in important life activities such as cell proliferation and differentiation, ontogeny, signal transduction, stem cell maintenance, and metabolism. It is closely related to the occurrence of a variety of major diseases. It participates in genome imprinting, X chromosome silencing, chromatin modification, and nuclear endoscopy. Transp...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888G01N33/68C12N15/113C12N15/11
CPCC12N15/113C12Q1/6888C12Q2600/124C12Q2600/158C12Q2600/178G01N33/68
Inventor 苗向阳黄万龙李嫒解领丽张秀秀
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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