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Construction method for Tp53-knockout animal model, and short peptides used therein

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 the problems of impure background, high cost, long preparation period, etc. Simple filtering process

Active Publication Date: 2018-10-23
BIOCYTOGEN PHARMACEUTICALS (BEIJING) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The first technical problem to be solved by the present invention is the problems of high cost, long preparation cycle and impure background in the preparation process of the p53 gene knockout rat model in the prior art, and further provides a low preparation cost, targeting Simple vector construction, short preparation cycle, and single background method for constructing an animal model for knocking out the Tp53 gene

Method used

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  • Construction method for Tp53-knockout animal model, and short peptides used therein
  • Construction method for Tp53-knockout animal model, and short peptides used therein
  • Construction method for Tp53-knockout animal model, and short peptides used therein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Design of Tp53 gene-specific TALEN sequence and construction of recognition module

[0038] 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:

[0039] LTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGRPALE;

[0040] The second short peptide has a sequence structure as shown in SEQ ID NO...

Embodiment 2

[0049] Example 2 Tp53 gene knockout transcriptional activator-like effector

[0050] 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.

[0051] 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:

[0052]CTGACCCCTGAGCAGGTGGTGGCCATCGCCAGCCACGACGGCGGCAAGCAGGCCCTGGAGACCGTGCAGAGGCTGCTGCCTGTGCTGTGCCAGGCCCACGGC...

Embodiment 3

[0055] Example 3 DNA vector comprising Tp53 gene knockout transcriptional activator-like effector

[0056] 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.

[0057] Specifically, the DNA vector is a DNA plasmid, and the DNA vector includes a first DNA vector and a second DNA vector.

[0058] 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 of is specifically:

[0059]

[0060] like 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: 6, the S...

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PUM

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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 Tp53 gene of a rat so as to rapidly acquire a Tp53-knockout rat. The invention also provides a method for knocking out the Tp53 gene and a construction method for a Tp53-knockout animal model. The acquired Tp53-knockout rat can be used for research related to tumors, including molecular biology study of tumors, research on neoplastic diseases, examination of carcinogenicity and the like, and has good application prospects.

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 Tp53 gene, and a method for constructing an animal model for knocking out Tp53 gene. Background technique [0002] Research on the mechanism of disease treatment and drug development for different human diseases has never stopped all over the world. However, due to the limitations of human ethics and technical realization, many studies cannot be directly carried out on humans, but need to be carried out on animals similar to humans. Experiment on the body. At present, the most commonly used in tumor biology research are transplanted tumor animal models and induced tumor animal models. However, the preparation process of transplanted tumor models is very demanding, requiring aseptic operation, and the shape of tumor cells, chromosome content, etc. have a great impact on th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/22C12N15/55C12N15/85A01K67/027
CPCA01K67/0278A01K2227/105C12N9/22C12N15/85C12N2810/00
Inventor 沈月雷张美玲白阳周小飞姚佳维苏幼红李冲杜吉超赵会珍
Owner BIOCYTOGEN PHARMACEUTICALS (BEIJING) CO LTD
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