Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Application of miRNA-sc-miR-145 to LC-PUFA synthesis of fish

A mirna-sc-mir-145, 1. mirna-sc-mir-145 technology, applied in the field of fish molecular marker preparation, can solve the problem of limiting the application of vegetable oil, reducing the LC-PUFA content of fish, and affecting aquaculture fish Growth and health problems, to achieve the effect of reducing the amount of use

Active Publication Date: 2020-07-28
SHANTOU UNIV
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the lack of LC-PUFA in vegetable oil, its substitution of fish oil not only affects the growth and health of aquaculture fish, but also significantly reduces the content of LC-PUFA in fish, which greatly limits the application of vegetable oil in compound feed.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of miRNA-sc-miR-145 to LC-PUFA synthesis of fish
  • Application of miRNA-sc-miR-145 to LC-PUFA synthesis of fish
  • Application of miRNA-sc-miR-145 to LC-PUFA synthesis of fish

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] To determine the expression level of sc-miR-145 in the liver tissue of the macula macula under feeding conditions of different fat sources.

[0023] The used macular bluefish used in the present invention comes from the wild fishing of Nan'ao Island, Shantou City, and is randomly divided into 2 groups after being domesticated in indoor salty fresh water (10ppt) for one month, with 3 tanks in each group and 20 fish in each tank, and throwing them in respectively. Feed a compound feed with vegetable oil or fish oil as the main fat source. After feeding for 8 weeks, the liver tissue of bluefish was collected, and the total RNA in the tissue was extracted using Trizol reagent provided by Roche Company. Six parallel samples of liver tissue in the same treatment group were mixed in equal amounts to obtain two mixed samples of FO and VO. Sent to Shenzhen BGI Institute for HiSeq small RNA high-throughput sequencing.

[0024] The results of HiSeq small RNA high-throughput seque...

Embodiment 2

[0027] Targeted regulation of sc-miR-145 on Hnf4α gene

[0028] In this example, the precursor sequence and its mature sequence of sc-miR-145 were cloned by chromosome walking technology, and the mature sequence is: GUCCAGUUUUCCCAGGAAUCCC. The stem-loop structure of the precursor sequence of sc-miR-145 is shown in figure 2 shown, from figure 2 It can be seen that the sc-miR-145 precursor sequence folds into a stable stem-loop structure, which belongs to the typical secondary structure of miRNA precursors and conforms to the structural characteristics of miRNA precursors. According to the principle of complementary pairing between the target gene and the seed sequence, nine consecutive base sequences that were completely complementary to the sc-miR-145 seed sequence were found on the 3'UTR of the Hnf4α gene.

[0029] In order to prove whether there is an interaction effect between the two, this example conducts a dual-luciferase reporter gene system experiment. The luciferas...

Embodiment 3

[0033] Regulatory effect of sc-miR-145 on Hnf4α and its downstream genes

[0034] Hnf4α has been proved to be a transcription factor that regulates the expression of key enzyme genes △4Fad, △6△5Fad and Elovl5 in the LC-PUFA synthesis of bluefish.

[0035] In order to verify the regulatory effect of sc-miR-145 on Hnf4α and its downstream genes, this example detected the protein and mRNA expression levels of Hnf4α and its downstream genes after overexpression or knockdown expression of sc-miR-145 in the hepatic cells of bluefish The change. The precursor sequence of sc-miR-145 and its stable inhibitor antagomir were synthesized in Guangzhou Ruibo Biotechnology Co., Ltd.

[0036] The results showed that after overexpression of sc-miR-145 precursor, the mRNA and protein expression levels of Hnf4α and downstream gene △4Fad were significantly decreased, and the mRNA levels of △6△5Fad and Elovl5 were also significantly down-regulated. On the contrary, after knocking down sc-miR-145...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to an application of miRNA-sc-miR-145 to LC-PUFA synthesis of fish. The fish is siganus canaliculatus, and the sequence table of miRNA-sc-miR-145 is shown as SEQID NO:1. The precursor sequence of the miRNA-sc-miR-145 is shown as SEQID NO:2. The DNA sequence of the precursor sequence of the miRNA-sc-miR-145 is shown as SEQID NO:3. Different fat source feeds are used for feeding the siganus canaliculatus, and response difference of miR-145 for different fat sources is verified, so that the miR-145 and a precursor pre-miR-145 thereof achieve important effects on synthesis and metabolism of long-chain polyunsaturated fatty acids. Molecular biology experiment verifies that the siganus canaliculatus miR-145 and the precursor pre-miR-145 thereof achieve important effects onLC-PUFA synthesis, and the knockdown expression of the siganus canaliculatus miR-145 and the precursor pre-miR-145 thereof can promote synthesis of LC-PUFA in fish bodies. The miR-145 can be used as amolecular marker associated with siganus canaliculatus long chain polyunsaturated fatty acid synthesis capacity, and has important practical application value in heredity assistance breeding.

Description

technical field [0001] The invention belongs to the technical field of fish molecular marker preparation, and specifically relates to the application of miRNA-sc-miR-145 in the synthesis of fish LC-PUFA. Background technique [0002] microRNA (miRNA) is a kind of single-stranded non-coding small RNA with a length of 18-25 nucleotides. Its transcription starts in the nucleus, and it is initially transcribed to form a typical stem-loop with a length of about 60-90 nucleotides. Structured pre-miRNA, then pre-miRNA is transported out of the nucleus, processed into mature miRNA by Dicer enzyme in the cytoplasm. miRNA can recognize the 3' non-coding region (3'UTR) of a specific target mRNA through sequence complementarity, degrade it or inhibit translation, and participate in gene post-transcriptional level regulation. [0003] Long-chain polyunsaturated fatty acids (LC-PUFA) are essential fatty acids (EFA) for vertebrates to maintain normal growth and development, mainly includi...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/113
CPCC12Q1/6888C12Q2600/178C12Q2600/158C12Q2600/124
Inventor 陈翠英王树启游翠红张媚胡宇
Owner SHANTOU UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com