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Long-chain non-coding RNA AAGNCR and application thereof

A long-chain non-coding, intramuscular fat technology, applied in the direction of DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., to achieve the reduction of marker genes, the reduction of lipid deposition in intramuscular fat cells, and the improvement of marker genes. Effect

Active Publication Date: 2021-03-30
INST OF ANIMAL HUSBANDRY & VETERINARY MEDICINE HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Through high-throughput sequencing in the early stage, this project screened a new lncRNA, named AAGNCR, which is highly expressed in intramuscular adipose tissue and continuously expressed in longissimus dorsi muscle of fat-type pig breeds (high intramuscular fat content) It is higher than lean pig breeds (low intramuscular fat content), suggesting that this lncRNA may be related to the trait of intramuscular fat content, and may be used as a marker of intramuscular fat content, but there is no report on the function of this lncRNA

Method used

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  • Long-chain non-coding RNA AAGNCR and application thereof
  • Long-chain non-coding RNA AAGNCR and application thereof
  • Long-chain non-coding RNA AAGNCR and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Expression of AAGNCR in Different Tissues of Pigs

[0022] 1. Test material

[0023] According to the standard sample collection plan, 3 Huainan pigs in the mid-fattening period (75 kg body weight) were randomly collected from Henan Xingrui Agriculture and Animal Husbandry Co., Ltd. All pigs were raised under the same conditions and growth environment, and the stomach, heart, liver, spleen, and lung were selected. , kidney, intestine, longissimus dorsi muscle, subcutaneous fat, intermuscular fat tissue samples.

[0024] 2. Test method

[0025] 2.1. Extraction of total RNA from different tissues of Huainan pigs

[0026] (1) Preparation: Use detergent to clean all mortars, grinding rods and sample spoons, soak in 3% hydrogen peroxide for more than 4 hours, and rinse with distilled water 3 times after washing. After drying, pack them in tin foil paper (to prevent contamination by RNase in the environment after taking them out), and then pack them in kraft pap...

Embodiment 2

[0040] Example 2: Differences in the expression of AAGNCR in pigs with different intramuscular fat content

[0041] 1. Test material

[0042] According to the standard sample collection plan, Huainan pigs (fat type pigs, high intramuscular fat content) and Landrace pigs (lean meat type pigs, low intramuscular fat content) were randomly collected from Henan Xingrui Agriculture and Animal Husbandry Co., Ltd. at 38 parities and 58 parities age, 7 days, and 130kg (Huainan pigs are about 300 days old, Landrace pigs are about 260 days old, and the growth speeds of the two pig breeds are different), and 10 pigs of each pig breed are collected at each stage. All pigs Only the feeding conditions are consistent with the growth environment.

[0043] 2. Test method

[0044] 2.1 Extract the total RNA from the longissimus dorsi muscle of Huainan pigs and Landrace pigs, the method is the same as in Example 1.

[0045] 2.2 Total RNA for reverse transcription PCR reaction

[0046] 2.3 Para...

Embodiment 3

[0054] Example 3: AAGNCR expression levels before and after adipocyte differentiation

[0055] 1. Test material

[0056] Cells: Porcine primary intramuscular adipocytes were isolated and cultured in this laboratory, frozen and stored in liquid nitrogen, and used after thawing after recovery.

[0057] Reagents: SYBRPremix Ex TaqTM (Tli RNaseH Plus) kit for real-time quantitative PCR (polymerase chain reaction) was purchased from TaKaRa Company. Real-time PCR specific primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd.

[0058] 2. Test method

[0059] 2.1 Isolation and culture of porcine primary intramuscular adipocytes

[0060] Three-day-old piglets were sacrificed under anesthesia, disinfected with Lysol solution and 75% alcohol, and separated about 5 g of longissimus dorsi muscle tissue. Rinse 3 times with PBS containing double antibody, remove connective tissue and blood vessels visible to the naked eye, and cut into 1-2mm 3 The minced meat was transf...

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Abstract

The invention belongs to the technical field of molecular biology, and particularly relates to a long-chain non-coding RNA AAGNCR and application thereof in prediction of intramuscular fat content ofpigs. The objective existence of AAGNCR is proved through RT-PCR, and meanwhile it is proved that AAGNCR has a promoting effect on intramuscular adipose cell lipid deposition. The AAGNCR siRNA sequence for inhibiting the expression of the AAGNCR sequence is synthesized by aiming at the AAGNCR sequence, and the sequence can obviously reduce the expression quantity of the AAGNCR in cells and has anobvious inhibition effect on the expression of the AAGNCR.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a long-chain non-coding RNA AAGNCR and its application in predicting pig intramuscular fat content. Background technique [0002] With the improvement of living standards, people's demand for high-quality pork is gradually increasing, and the price of products related to local pig breeds in my country is significantly higher than that of imported breeds. The essence of high-quality pork is small muscle fiber diameter and high intramuscular fat content. Intramuscular fat content is one of the key factors affecting meat quality, affecting tenderness, flavor and juiciness of meat. Intramuscular fat content is regulated by many factors, such as genetics, environment, nutritional level, etc., and the influence of genetics is the most critical. Intramuscular fat content is a quality trait, which is regulated by multiple genes with minor effects, and its heritabi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12Q1/6888
CPCC12N15/113C12Q1/6888C12Q2600/124C12Q2600/158C12Q2600/178C12N2310/14Y02P60/87
Inventor 王璟陈俊峰滑留帅邢宝松卢清侠郭红霞马强张华张家庆任巧玲孙加节潘传英
Owner INST OF ANIMAL HUSBANDRY & VETERINARY MEDICINE HENAN ACAD OF AGRI SCI
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