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Construction method for Il2rg-knockout animal model, and short peptides used therein

A technology for constructing methods and animal models, applied in biochemical equipment and methods, chemical instruments and methods, peptides, etc., can solve the problems of high preparation cost, complex screening process, and high cytotoxicity of knockout rat models, and achieve improved preparation. Effects of low success rate, low off-target rate, and low cytotoxicity

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

[0009] The first technical problem to be solved by the present invention is the high cost of preparation of the Il2rg gene knockout rat model in the prior art, the complex construction and screening process of the targeting vector, high cytotoxicity, high off-target rate, and limited application , and further provide a method for constructing and knocking out the Il2rg gene with low preparation cost, simple targeting vector construction and screening process, low cytotoxicity, low off-target rate, and wide application

Method used

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  • Construction method for Il2rg-knockout animal model, and short peptides used therein
  • Construction method for Il2rg-knockout animal model, and short peptides used therein
  • Construction method for Il2rg-knockout animal model, and short peptides used therein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Design of Il2rg Gene-Specific TALEN Sequence and Construction of Recognition Module

[0044] 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:

[0045] LTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGRPALE;

[0046] The second short peptide has a sequence structure as shown in SEQ ID N...

Embodiment 2

[0055] Example 2 Il2rg gene knockout transcriptional activator-like effector

[0056] 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.

[0057] 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:

[0058]CTGACCCCTGAGCAGGTGGTGGCCATCGCCAGCAACAACGGCGGCAAGCAGGCCCTGGAGACCGTGCAGAGGCTGCTGCCTGTGCTGTGCCAGGCCCACGG...

Embodiment 3

[0061] Example 3 DNA vector comprising Il2rg gene knockout transcriptional activator-like effector

[0062] 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.

[0063] Specifically, the DNA vector is a DNA plasmid, including a first DNA vector and a second DNA vector.

[0064] like 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:

[0065]

[0066] 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 SEQ ID NO: 6 The seq...

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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 Il2rg gene of a rat so as to rapidly acquire an Il2rg-knockout rat. The invention also provides a method for knocking out the Il2rg gene and a construction method for an Il2rg-knockout animal model. The acquired Il2rg-knockout rat can be used for research on allotransplantation of tumors, development of humanized rats and research on autoimmune and infectious diseases, 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 Il2rg gene, and a method for constructing an animal model for knocking out Il2rg gene. Background technique [0002] The interleukin-2 receptor gamma chain (IL2Rγc) encoded by the IL2RG gene is a receptor component of various interleukin signaling pathways, mainly responsible for mediating interleukin family factors IL-2, IL-4, IL-7, IL -9, IL-15 and IL-21 signal transduction. Studies have found that human IL2RG gene mutations can cause severe combined immunodeficiency (X-linked severe combined immunodeficiency, X-SCID), which seriously affects the differentiation and maturation of T cells and NK cells in the body as well as B cells. normal function, resulting in a severe loss of immunity. As a kind of cellular and humoral immune deficiency, X-SCID accounts for about 50-...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/00C12N15/11C12N15/63A01K67/027
CPCA01K67/0276A01K2207/15A01K2217/075A01K2227/105A01K2267/03C07K14/00C07K14/7155C12N15/63C12N2800/80
Inventor 沈月雷张美玲白阳周小飞苏幼红姚佳维杜吉超赵会珍李冲
Owner BIOCYTOGEN PHARMACEUTICALS (BEIJING) CO LTD
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