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A kind of preparation method of gene editing pig with "imitation Belgian blue cow" mstn genotype

A technology imitating Belgian blue cattle and genotypes, applied in the field of gene-edited pig preparation

Active Publication Date: 2017-01-18
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

Sequencing of the Belgian blue cattle bred by natural selection revealed that the deletion of the conserved biologically active region 11bp in exon 3 of the MSTN gene caused the double muscle phenomenon

Method used

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  • A kind of preparation method of gene editing pig with "imitation Belgian blue cow" mstn genotype
  • A kind of preparation method of gene editing pig with "imitation Belgian blue cow" mstn genotype
  • A kind of preparation method of gene editing pig with "imitation Belgian blue cow" mstn genotype

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1. Preparation of gene-edited pigs of MSTN genotype "Imitation Belgian Blue Cattle"

[0038] 1. Preparation of MSTN genotype cells containing "imitation Belgian blue cow"

[0039] 1. Obtaining porcine embryonic fibroblast (PEF) in vitro

[0040] 35 embryos of different breeds of pigs ( figure 1 ), remove the head, tail, limbs, viscera and bones of the fetus, and clean up the blood. Continue to cut the fetus with elbow ophthalmic scissors for 30 minutes to ensure that it is fully shredded. Pipette the shredded fetal tissue into a 15mL centrifuge tube with the blue tip of the scissors, add 5mL of full medium, and remove the above solution after natural sedimentation for several minutes. Add a few drops of FBS to the lower tissue block, suck it out with a 15cm glass Pasteur tube bent at 1cm from the tip, spread it in two T75 culture flasks, place the bottom of the flask upwards, and add 15mL full culture solution on the opposite side, After 6-8 hours, turn the c...

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Abstract

The invention discloses a method for preparing an 'imitated Belgian blue cattle' myostatin (MSTN) genetype gene editing pig. By adopting the method provided by the invention, a targeted area in an MSTN gene exon 3 of a target porcine genome is subjected to genome editing, so that the exon 3 forms a terminator codon in advance to terminate expression, so as to obtain the 'imitated Belgian blue cattle' MSTN genetype gene editing pig. Compared with the other mutation types (not forming frameshift mutation or terminator codon in advance) of gene editing, by adopting the 'imitated Belgian blue cattle' MSTN genetype gene editing pig prepared by the method, the terminator codon can be formed in advance, and the method is more effective to modification of the MSTN gene.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preparing a gene-edited pig of the "imitation Belgian blue cow" MSTN genotype. Background technique [0002] Myostatin (MSTN) is a negative regulator of muscle growth, and its mutation can cause myocyte hyperplasia and hypertrophy, resulting in an increase in muscle mass. In addition, because it is a specific factor for muscle development, the genetic engineering operation on it will not affect other tissues, so it has broad application prospects and commercial value in many aspects such as agriculture, fishery, animal husbandry and medicine. [0003] my country is the largest pork consumption country in the world, and many breeders are devoted to the breeding of pig breeds with high lean meat percentage. Sequencing of the Belgian blue cattle bred by natural selection revealed that in exon 3 of the MSTN gene, the deletion of the conserved biologically active region 11bp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/073C12N15/85C12N15/877A01K67/027
Inventor 李奎牟玉莲吴添文阮进学黄雷魏景亮
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI