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NOD-genetic background neutropenic humanized mouse model preparation method

A neutrophil and mouse model technology, applied in the field of genetic engineering and genetic modification, to achieve highly original effects

Active Publication Date: 2018-07-20
XINXIANG MEDICAL UNIV
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Problems solved by technology

[0004] NOD mice are the non-obese diabetic strain (NOD) and non-obese normal strain (nod) isolated from cataract-susceptible substrains during the 6th generation of inbred breeding of outbred strain Jcl: JCR mice by Japanese scholars. The most widely used mouse strains in diabetes research and establishment of humanized models. At present, some classic humanized mouse models such as NOG and NSG are produced in the genetic background of NOD mice; neutrophils are An important member of immune cells, which is crucial to the survival of mice, there is no report on the successful production of humanized mouse models of neutrophil deficiency in the world; therefore, mice with NOD genetic background The production of a humanized mouse model of neutrophil deficiency will provide a better mouse model for the study of such mice in xenotransplantation, and also open a new perspective for the study of humanized mice

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  • NOD-genetic background neutropenic humanized mouse model preparation method

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Embodiment 1

[0039] The preparation method of the humanized mouse model of neutrophil deficiency in the NOD genetic background in this embodiment comprises the following steps:

[0040] 1. Determine the three specific target sites Gfi1-sgRNA1, Gfi1-sgRNA2, and Gfi1-sgRNA3 of the gene Gfi1 (Gene ID: 103170) to be knocked out in NOD mice.

[0041]Find the mouse Gfi1 gene DNA sequence in the mouse genome database ensembl (http: / / asia.ensembl.org), and then use the online design software CRISPOR (http: / / crispor.tefor.net / crispor.cgi) to determine the Three specific sites were selected in exon6 (exon ID: ENSMUSE00001072804) of the mouse Gfi1 gene as the target sequence of the sgRNA. The three target sequences are:

[0042] Gfi1-sgRNA1: 5'-AGCTTTGACTGTAAGATCTG TGG-3' (as shown in SEQ ID NO.1);

[0043] Gfi1-sgRNA2: 5'-GTACTGACAGGGATAGGGCC GGG-3' (as shown in SEQ ID NO.2);

[0044] Gfi1-sgRNA3: 5'-TGCTCATTCACTCGGACACC CGG-3' (as shown in SEQ ID NO.3).

[0045] 2. Obtain 3 sgRNA mRNAs.

[0046...

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Abstract

The invention relates to a preparation method of an NOD-genetic background neutropenic humanized mouse model, and belongs to the technical field of genetic engineering and genetic modification. The preparation method comprises the step of knocking out a Gfi1 gene from an NOD mouse to obtain the NOD-genetic background neutropenic humanized mouse model. The invention achieves specific knockout of the key gene Gfi1, which controls mouse neutrophil development, from the NOD-genetic background mouse model through a CRISPR / Cas9 system for the first time, and the novel NOD-genetic background neutropenic humanized mouse animal model is obtained. The mouse model has the potential of accepting heterotransplantation and provides a new concept for development of a brand-new humanized mouse.

Description

technical field [0001] The invention relates to a preparation method of a neutrophil-deficient humanized mouse model with NOD genetic background, and belongs to the technical field of genetic engineering and genetic modification. Background technique [0002] Humanized mouse model refers to a mouse model with human functional genes, cells or tissues. In order to better rebuild the human immune system in humanized mice, it is necessary to remove the mouse's own immune system first. Many classic humanized mouse models have been obtained by previous studies, such as NOG and NSG mice with complete deletion of T cells, B cells and natural killer cells. [0003] The human immune system is a very important and complex network system, which is inextricably linked to the occurrence of almost all human diseases. In order to deeply study the relationship between the human immune system and diseases, it is necessary to use various animal models To simulate some special diseases, due to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/90C12N15/89C12N9/22A01K67/027
CPCA01K67/0275A01K2227/105A01K2267/03C12N9/22C12N15/89C12N15/907
Inventor 梁银明张黎琛卢燎勋黄蓉晁天柱郑前前罗静谷妍蓉
Owner XINXIANG MEDICAL UNIV
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