A method of constructing a zygote of a microRNA-overexpression-based animal with liver cancer

An overexpression and fertilized egg technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, the use of vectors to introduce foreign genetic material, etc., can solve the problems of high operator requirements, inability to pass generations, cumbersome operation of liver cancer animal models, etc.

Inactive Publication Date: 2015-07-08
HARBIN MEDICAL UNIVERSITY
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Problems solved by technology

However, the above animal models of liver cancer have disadvantages such as cumbersome operation, inability to pass passage, and high requirements for operators.

Method used

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  • A method of constructing a zygote of a microRNA-overexpression-based animal with liver cancer
  • A method of constructing a zygote of a microRNA-overexpression-based animal with liver cancer
  • A method of constructing a zygote of a microRNA-overexpression-based animal with liver cancer

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Embodiment Construction

[0020] The present invention will be further described through experiments and examples below. It should be understood that these examples are only for the purpose of illustration, and in no way limit the protection scope of the present invention.

[0021] Concrete technical scheme of the present invention is:

[0022] 1. Construction of CAGG-pre-miR-483 plasmid

[0023] Select the mouse miR-483 sequence (Gene ID: 723874) on NCBI and its upstream and downstream flanking sequences of 150 bp, and design cloning primers containing EcoR I restriction sites. The primers are: miR483-CL-F:ggtc GAATTC cactcctgcacggaggg, miR483-CL-R: caggt GAATTC cctgtacttggggcccg (the underline indicates the enzyme cleavage site). Using the genome as a template, its sequence was amplified by PCR, and the product size was 237bp. The PCR product obtained by EcoR I single enzyme digestion and the carrier pCAGGs containing the promoter CAGG were 3h, CIP dephosphorylated the vector for 1h, and 5μg produ...

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Abstract

The invention relates to a method of constructing a zygote of a microRNA-overexpression-based animal with liver cancer, and belongs to the technical field of biology. The method firstly constructs a CAGG-pre-miR-483 plasmid, and then constructs the zygote of an miR-483-overexpressed transgenic mouse. A genetic change mouse model based on stable microRNA overexpression is built. The model is achieved by integrating an exogenous nucleic acid sequence into mouse genome. The building of the model provides a reliable platform for vector function research of microRNAs in liver cancer.

Description

technical field [0001] The invention specifically relates to a method for establishing fertilized eggs of liver cancer animals based on microRNA overexpression, and belongs to the field of biotechnology. Background technique [0002] Hepatocellular carcinoma (HCC, referred to as "liver cancer") is the most common liver tumor. my country is an area with a high incidence of hepatitis B and alcoholic liver disease, and a high incidence and mortality rate of liver cancer. Although surgery, radiotherapy and chemotherapy have been carried out clinically, the treatment effect is poor due to the high degree of malignancy of the tumor itself. Therefore, establishing a good liver cancer animal model is crucial for new drug development and translational medical research. [0003] From the perspective of the species of model animals, mice are widely used due to their small footprint, low cost, fast reproduction, high homology with human genome and similar pathological characteristics....

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C12N15/113C12N5/10
Inventor 马宁高旭刘彦虹张艳芬颜炳柱邱前义周凌云王磊
Owner HARBIN MEDICAL UNIVERSITY
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