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Application of ZC3H12b gene or protein and establishment method of hepatic disease animal model

A gene and protein technology, applied in biochemical equipment and methods, disease diagnosis, peptide/protein components, etc., can solve problems such as knockout animal models of liver diseases

Pending Publication Date: 2021-07-16
SHANGHAI OCEAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, as of the filing date of this patent, there is no report on the relationship between Zc3h12b and liver disease, especially ICC and its related fatty liver and liver cancer, and there is no report on the animal model of liver disease obtained by knocking out Zc3h12b in mice or any other animals

Method used

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  • Application of ZC3H12b gene or protein and establishment method of hepatic disease animal model
  • Application of ZC3H12b gene or protein and establishment method of hepatic disease animal model
  • Application of ZC3H12b gene or protein and establishment method of hepatic disease animal model

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Embodiment 1

[0066] 1. Zc3h12b gene knockout and identification

[0067] Using the CRISPR / Cas9 system, the zc3h12b gene was knocked out by microinjecting the one-cell stage fertilized eggs of medaka. The specific method is as follows:

[0068] (1) Obtain the medaka (Japanese medaka HdrR, Oryzias latipes) zc3h12b gene sequence through the enseble website (http: / / asia.ensembl.org / index.html). On the crisprscan website (http: / / crisprscan.org / ), according to the medaka zc3h12b gene sequence, search for the targeted knockout sites of the CRISPR / CAS9 editing system, and at the same time search through the BLAST comparison of the medaka transcriptome, select no Any other candidate sites for non-specific binding are predicted. The target site is chosen to be near the first ATG after the promoter. Add the T7 promoter sequence before the target sequence, and add the gRNA scaffold sequence at the back end. At the same time, in order to ensure the efficiency of the T7 promoter, the first two bases ...

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Abstract

The invention relates to application of a ZC3H12b gene or protein and a method for establishing a hepatic disease animal model. According to the invention, a gene editing technology is utilized, the zc3h12b gene of oryzias latipes is knocked out in a targeted mode, the oryzias latipes with Zc3h12b protein product deficiency are established, all the oryzias latipes show different degrees of hepatobiliary duct hyperplasia and fusion, hepatocyte steatosis, fibrosis and other hepatic lesions, the obvious fatty liver appears along with the month-old growth, local cyst necrosis occurs, lymphocyte infiltration is obvious in the hepatic sinusoid, the number of macrophages is increased abnormally, positive cells of human tumor markers CK19, SMA and GPC3 are detected, and it is suggested that ZC3H12b can serve as a therapeutic target and a biomarker of intrahepatic biliary cystadenoma, intrahepatic biliary cystadenocarcinoma or the related fatty liver or liver cancer. The zc3h12b-deficient oryzias latipes can be used as an animal model for researching the intrahepatic biliary cystadenoma, the intrahepatic biliary cystadenocarcinoma or the lesion process of the intrahepatic biliary cystadenoma and the intrahepatic biliary cystadenocarcinoma.

Description

technical field [0001] The present invention relates to the field of biomedical technology, in particular to the establishment of new functions of genes or proteins and animal models of related diseases, specifically, the use of ZC3H12b genes or proteins and the establishment of animal models for simulating liver diseases. Background technique [0002] Liver diseases are divided into non-neoplastic and neoplastic liver diseases, but non-neoplastic liver diseases (common pathogenic infections such as hepatitis viruses or parasites, chemical or alcohol-induced liver injury, and genetic abnormalities such as cholelithiasis and bile duct deformity) , because the microenvironment where the liver cells, bile duct epithelial cells and stem cells in the liver sinusoids are constantly stimulated by the bile secreted by the bile duct epithelium, the intrinsic macrophages (Kupffer cells, KC) of the liver sinusoids and other lymphocytes that regulate the autoimmune system The influence ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61P35/00A61P1/16A61K49/00C12Q1/6886C12Q1/6883G01N33/68G01N33/574
CPCA61K45/00A61P35/00A61P1/16A61K49/0008C12Q1/6886C12Q1/6883G01N33/6893G01N33/57484G01N33/57438C12Q2600/136C12Q2600/158G01N2800/085A61K49/00G01N33/574G01N33/68A61K38/00C12N15/85
Inventor 关桂君常宇阳
Owner SHANGHAI OCEAN UNIV
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