A method for constructing a spontaneous anemia mouse model with leukocyte and thrombocytosis

A technology of thrombocytosis and mouse models, applied in other methods of inserting foreign genetic materials, chemical instruments and methods, biochemical equipment and methods, etc.

Active Publication Date: 2021-08-31
ZHEJIANG CHINESE MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Clinically reported anemia combined with leukocyte hyperplasia accounts for a large proportion, and the anemia disease mouse model of leukocyte and thrombocytosis obtained by site-directed mutation of STAT3 has not been reported so far

Method used

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  • A method for constructing a spontaneous anemia mouse model with leukocyte and thrombocytosis
  • A method for constructing a spontaneous anemia mouse model with leukocyte and thrombocytosis
  • A method for constructing a spontaneous anemia mouse model with leukocyte and thrombocytosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Construction of spontaneous anemia model mice (i.e. STAT3-K177 / 180R mutant mice) accompanied by leukocyte and thrombocytosis

[0029] 1. Crispr / Cas9 site design: use CRISPR / Cas9 technology to find the target sequence for the gene structure of the target gene, design guide-RNA for this site according to the target sequence information, and after activity detection, guide-RNA with activity will be RNA and Cas9 were transcribed into RNA in vitro, and guide-RNA, Cas9 mRNA, and oligo donor DNA for recombination were injected into fertilized eggs by microinjection, and positive knockout mice were screened from the offspring.

[0030] According to the above analysis, a guide-RNA (SEQ ID NO.1) was designed for the upstream and downstream sequences of exon No. 6 (82bp), where the mouse mutation site is located. Site 1: The exon6 target sequence is as follows: (SEQ ID NO.4, the underline is the mutation site, the uppercase is exon, the lowercase is intron)

[0031] SEQ...

Embodiment 2

[0050] Example 2: Index detection of STAT3-K177 / 180R mutant mice

[0051] 1. Observation of the phenotype of the mutant mice: observe the surviving STAT3-K177 / 180R mutant homozygous mice, and find that the homozygous mice grow slowly and become smaller ( Figure 4 Middle A), 12 homozygous, 12 heterozygous and wild-type mice were extracted at the age of 12 weeks, and the weight statistics were carried out, which showed that the weight loss of the homozygous mice was statistically significant ( Figure 4 Middle B), after continuous observation of the growth difference between homozygous mice and wild-type mice, it was found that the hair of homozygous mice began to turn yellow and fall off at the later stage of growth, and some hairs turned white, and some even had skin ulcers ( Figure 4 Middle C).

[0052] 2. Spleen index statistics of mutant mice: 12-week-old homozygous (mut / mut) mice and wild-type (wt / wt) mice were randomly selected for dissection, and the spleen of the hom...

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Abstract

The invention discloses a method for constructing a spontaneous anemia mouse model with leukocytes and thrombocytosis. The method is to mutate the 177th and 180th lysines of the mouse STAT3 protein into arginines. In the present invention, by performing site-directed mutation on the STAT3 gene, the 177th and 180th lysines of STAT3 are site-directedly mutated into arginine, and it is found that the mice show that white blood cells including lymphocytes, granulocytes, and monocytes are all increased, and platelets Anemia models with elevated but significantly reduced red blood cells and hemoglobin are also shown. The mouse model of the disease established in the present invention is of great significance for the study of the occurrence and development mechanism of anemia accompanied by the increase of white blood cells and platelets and the development of anti-drugs.

Description

[0001] (1) Technical field [0002] The invention relates to a method for constructing a spontaneous anemia mouse model with leukocytes and thrombocytosis. [0003] (2) Background technology [0004] Anemia is a common blood system disease, which refers to a disease in which the number of red blood cells contained in the peripheral blood of the patient's body is significantly reduced, and the hemoglobin cell value and red blood cell value are lower than normal levels. Clinically, there are many pathogenic factors of anemia, such as immune system disorders, long-term malnutrition, or cellular lesions in the body itself. [0005] There are many types of anemia, which can be divided into two types according to physiological characteristics, anemia caused by hematopoietic system and anemia caused by non-hematopoietic system. Anemia caused by hematopoietic system diseases accounts for about 40%-45%, mainly megaloblastic anemia, followed by acute leukemia, aplastic anemia, chronic m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/90A01K67/027
CPCA01K67/0276A01K2217/075A01K2227/106A01K2267/0381C07K14/705C12N15/113C12N15/902C12N2310/10C12N2310/20
Inventor 徐莉季巾君秦樾范永升杜羽
Owner ZHEJIANG CHINESE MEDICAL UNIVERSITY
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