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Method for constructing animal model of apoe gene knockout and short peptide thereof

A technology for constructing methods and animal models, applied in the fields of botanical equipment and methods, biochemical equipment and methods, genetic engineering, etc., can solve problems such as high blood lipids, and achieve the effect of improving reliability, rapid editing and genetic modification

Active Publication Date: 2020-10-09
BIOCYTOGEN PHARMACEUTICALS (BEIJING) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The first technical problem to be solved by the present invention is that in the preparation of the Apoe gene knockout rat model in the prior art, there is still the possibility of restoring the expression of Apoe protein, and it is impossible to make the rats show the characteristics of hyperlipidemia in the early adulthood problem, and then provide a method for constructing a rat model of knocking out the Apoe gene, which has more reliable results and can show the characteristics of hyperlipidemia in the early adulthood of rats

Method used

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  • Method for constructing animal model of apoe gene knockout and short peptide thereof
  • Method for constructing animal model of apoe gene knockout and short peptide thereof
  • Method for constructing animal model of apoe gene knockout and short peptide thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The design of embodiment 1 Apoe gene-specific TALEN sequence and the construction of recognition module

[0037]A pair of short peptides in this embodiment, the short peptides include a first short peptide and a second short peptide; the first short peptide has a sequence structure as shown in SEQ ID NO: 1, and the SEQ ID NO: The sequence structure of 1 is specifically:

[0038] LTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGRPALE;

[0039] The second short peptide has a sequence structure as shown in...

Embodiment 2

[0048] Example 2 Apoe gene knockout transcriptional activator-like effector

[0049] In order to encode the first short peptide and the second short peptide described in Example 1, a pair of transcriptional activator-like effectors is constructed in this embodiment, and the transcriptional activator-like effectors include the first transcriptional activator-like effector and the first transcriptional activator-like effector The second transcription activator-like effector; the first transcription activator-like effector and the second transcription activator-like effector are respectively transcribed and translated to obtain the first short peptide described in Example 1 and the second short peptide.

[0050] The first transcriptional activator-like effector has a sequence structure as shown in SEQ ID NO: 3, and the sequence structure of SEQ ID NO: 3 is specifically:

[0051] CTGACCCCTGAGCAGGTGGTGGCCATCGCCAGCAACGGCGGCGGCAAGCAGGCCCTGGAGACCGTGCAGAGGCTGCTGCCTGTGCTGTGCCAGGCCCACGG...

Embodiment 3

[0054] Embodiment 3 comprises the DNA carrier of Apoe gene knockout transcriptional activator-like effector

[0055] The DNA vector of this example contains the transcriptional activator-like effector described in Example 2; as a specific implementation of this example, the DNA vector is a DNA vector that can encode a FokI nucleic acid.

[0056] Specifically, the DNA vector is a DNA plasmid, and the DNA vector includes a first DNA vector and a second DNA vector.

[0057] Such as figure 1 Shown is a schematic diagram of the construction of the first DNA vector, the first DNA vector can recognize the L sequence described in Example 1, and has a sequence structure as shown in SEQ ID NO: 5, the SEQ ID NO: 5 The sequence structure is specifically:

[0058]

[0059] Such as figure 2 Shown is a schematic diagram of the construction of the second DNA vector, the second DNA vector can recognize the R sequence described in Example 1, and has a sequence structure as shown in SEQ ID NO...

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Abstract

The invention provides a pair of short peptides, a pair of transcription activator-like effectors and a pair of DNA vectors. The short peptides of the invention can realize accurate and efficient targeting of the Apoe gene of a rat so as to rapidly acquire an Apoe-knockout rat. The invention also provides a method for knocking out the Apoe gene and a construction method for an Apoe-knockout animalmodel. The method for knocking out the Apoe gene is more reliable in gene knockout results and allows a rat to present the characteristic of hyperlipidemia in the early adult stage.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a pair of short peptides, a pair of transcription activator-like effectors, a method for knocking out the Apoe gene, and a method for constructing an animal model for knocking out the Apoe gene. Background technique [0002] ApoE protein is an apolipoprotein that exists in very low-density lipoprotein, chylomicrons, intermediate-density lipoprotein and high-density lipoprotein, and is mainly synthesized in liver and brain tissue, and participates in the transformation and metabolism of lipoproteins. Important ligands in lipid metabolism. Studies have shown that ApoE gene defect or gene variation is related to cardiovascular disease, Alzheimer's disease, age-related macular degeneration and other diseases, and is the main gene of hypercholesterolemia and atherosclerosis. [0003] Research on the mechanism of disease treatment and drug development for different human diseas...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/22C12N15/55C12N15/85A01K67/027
CPCA01K67/0278A01K2217/075A01K2227/105A01K2267/03C12N9/22C12N15/85
Inventor 沈月雷姚佳维张美玲白阳周小飞苏幼红余巧玲杜吉超赵会珍
Owner BIOCYTOGEN PHARMACEUTICALS (BEIJING) CO LTD
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