Marker Lnc-482286 related to porcine skeletal muscle satellite cell proliferation, and application of marker Lnc-482286

A technology of lnc-482286 and satellite cells, applied in the field of molecular biology, can solve the problem of less research on porcine tissue-specific LncRNA

Active Publication Date: 2021-09-07
SHANXI AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on the regulation of LncRNA is mostly focused on model animals such as mice, and there are few studies on porcine tissue-specific LncRNA

Method used

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  • Marker Lnc-482286 related to porcine skeletal muscle satellite cell proliferation, and application of marker Lnc-482286
  • Marker Lnc-482286 related to porcine skeletal muscle satellite cell proliferation, and application of marker Lnc-482286
  • Marker Lnc-482286 related to porcine skeletal muscle satellite cell proliferation, and application of marker Lnc-482286

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Validation of porcine long non-coding RNA Lnc-482286

[0024] Samples: Longissimus dorsi muscle tissue from Jinfen white pigs that were 1 day old was collected, wrapped in tinfoil paper, quickly frozen in liquid nitrogen, and stored at -80°C.

[0025] Reagent: Reagent was purchased from Life Technologies; PrimeScript RT reagentKit with gDNA Eraser and DNA marker were purchased from TaKaRa; 2×Es Taq Master Mix (Dye) was purchased from Kangwei Century Biotechnology Co., Ltd.; 50×TAE buffer, nucleic acid dye, Triton X-100 and DAPI staining solution were purchased from Suleibao Company; agarose was purchased from Shanghai Sangon Bioengineering Co., Ltd.

[0026] 1) Extraction of total RNA

[0027] Take out the longissimus dorsi muscle tissue of Jinfen white pig 1 day old from the -80°C refrigerator, add 1mL Trizol reagent, grind it fully with a tissue homogenizer, grind and pulverize it, let it stand for 10min, and fully lyse it at 4°C, 12000r· min -1 Centrifuge for 10...

Embodiment 2

[0037] Temporal expression properties of Lnc-482286

[0038] Samples: Longissimus dorsi muscle tissues of Jinfen white pigs were collected for 1 day, 90 days and 180 days respectively, wrapped in tinfoil paper, frozen in liquid nitrogen immediately, and stored at -80°C.

[0039] Reagent: Reagent was purchased from Life Technologies; PrimeScript RT reagent Kit with gDNA Eraser and 2×SYBR Premix Ex Taq II were purchased from TaKaRa.

[0040] According to the sequence (SEQ ID NO:1) of the Lnc-482286 transcript, the Lnc-482286 quantitative primer pair was designed by Oligo7 software, as follows:

[0041] Quantitative primer pair for Lnc-482286:

[0042] Upstream primer: 5'-AACAGGTGAGCAGGATGCGAAG-3' (SEQ ID NO: 4)

[0043] Downstream primer: 5'-GATCACAGTGCCCGTCCTTCAG-3' (SEQ ID NO:5)

[0044] Quantitative primer pairs for 18S rRNA:

[0045] Upstream primer: 5'-CCCACGGAATCGAGAAAGAG-3' (SEQ ID NO: 6)

[0046] Downstream primer: 5'-TTGACGGAAGGGCACCA-3' (SEQ ID NO: 7)

[0047] ...

Embodiment 3

[0051] The role of Lnc-482286 in the proliferation of porcine skeletal muscle satellite cells

[0052] Cells: Porcine skeletal muscle satellite cells were isolated and cultured in the laboratory.

[0053] Reagents: fetal bovine serum (FBS), DME / F12 were purchased from Gibco; penicillin-streptomycin mixture, glycine, 4% paraformaldehyde were purchased from Solebol; Triton X-100 was purchased from Sigma; 0.25% pancreatic Enzymes and PBS buffer were purchased from Hyclon; EdU proliferation detection kit and transfection reagent ribo FECT TM CP was purchased from Guangzhou Ruibo Biotechnology Co., Ltd.

[0054] For the Lnc-482286 sequence, synthesize an siRNA sequence (Smart Silencer) that inhibits its expression. SmartSilencer is a mixture, and the specific sequence is:

[0055] 5'-CGACATGCTCTCTTGGGTCT-3' (SEQ ID NO: 8)

[0056] 5'-CACGTGCCAGGAGACTTGTA-3' (SEQ ID NO:9)

[0057] 5'-CAGAACAGGTGAGCAGGAT-3' (SEQ ID NO: 10)

[0058] 5'-TCTTGTTTCCATGGTGACT-3' (SEQ ID NO: 11)

[...

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Abstract

The invention discloses a long non-coding RNA marker Lnc-482286 related to porcine skeletal muscle satellite cell proliferation, and application of the long non-coding RNA marker Lnc-482286, and belongs to the technical field of molecular biology. The long non-coding RNA marker is Lnc-482286, and the nucleotide sequence of the long non-coding RNA marker Lnc-482286 is shown as SEQ ID NO.1. The marker Lnc-482286 provided by the invention is highly expressed in porcine muscle tissues, expression of the Lnc-482286 in the muscle tissue of the Jinfen white pig is proved to be in a trend of firstly increasing and then decreasing along with the increase of the day age, and the Lnc-482286 plays a significant up-regulation role in the process of the porcine skeletal muscle satellite cell proliferation. The proliferation condition of the porcine skeletal muscle satellite cells can be identified by detecting the expression of the Lnc-482286, the porcine skeletal muscle satellite cell proliferation can be accelerated by improving the expression of the Lnc-482286, and a new thought and method is provided for clinical research on the growth and development of the porcine skeletal muscle.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a marker Lnc-482286 related to the proliferation of porcine skeletal muscle satellite cells and its application. Background technique [0002] The growth and development of skeletal muscle is inseparable from the proliferation and differentiation of myoblasts, and maintaining a dynamic balance between cell proliferation and differentiation is crucial to the growth and development of animal organisms. Skeletal muscle satellite cells are a kind of muscle-derived stem cells, which are located between the sarcolemma and the extracellular matrix of muscle fibers, that is, the basement membrane. The cells are spindle-shaped or spindle-shaped, and have the potential for proliferation and differentiation. Under normal conditions in the body, skeletal muscle satellite cells are in an inactive, quiescent state and fuse with established muscle fibers to maintain the stem cell pool...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N5/077C12N15/113
CPCC12Q1/6888C12N5/0659C12Q2600/178C12Q2600/158
Inventor 杨帅秦本源路畅曹果清吴怡琦李文霞杨阳蔡春波赵燕高鹏飞郭晓红李步高
Owner SHANXI AGRI UNIV
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