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Application of a mutant zebrafish in the preparation of an animal model of myelodysplastic syndrome

A technology of bone marrow dysplasia and animal models, applied in the direction of stably introducing foreign DNA into chromosomes, using microinjection, biochemical equipment and methods, etc., can solve the problem of unclear specific mechanism of leukemia

Active Publication Date: 2021-05-14
SOUTH CHINA UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, how ASXL1 specifically affects hematopoietic development and the specific mechanism of leukemia after mutation is still unclear.

Method used

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  • Application of a mutant zebrafish in the preparation of an animal model of myelodysplastic syndrome
  • Application of a mutant zebrafish in the preparation of an animal model of myelodysplastic syndrome
  • Application of a mutant zebrafish in the preparation of an animal model of myelodysplastic syndrome

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Embodiment 1

[0049] (1) Zebrafish farming

[0050] Zebrafish were cultured as described in the literature (Westerfield M: The zebrafish: guide for the laboratory use of zebrafish (Brachdanio rerio). Edition by Eugene, OR, M. Westerfield, 1993).

[0051] The following strains are used in the present invention: AB wild-type zebrafish, asxl1 mutant zebrafish and asxl1 mutant zebrafish with Tg(lyz:DsRED2) transgenic background; wherein,

[0052] Transgenic zebrafish Tg(lyz:DsRED2) reference (Hall C, Flores MV, Storm T, Crosier K, Crosier P. The zebrafish lysozyme C promoter drives myeloid-specific expression in transgenic fish. BMC Dev Biol 2007; 7:42.) was obtained (Tg(lyz:DsRED2) is the same as lysC::DsRED2 in ref.). Asxl1 mutant zebrafish with Tg(lyz:DsRED2) transgenic background was obtained by crossing asxl1 mutant zebrafish with transgenic zebrafish Tg(lyz:DsRED2).

[0053] (2)CRISPR / cas9

[0054] Mix gRNA and Cas9mRNA together so that the final concentration is gRNA (50-100ng / L), Cas...

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Abstract

The invention discloses the application of a mutant zebrafish in preparing an animal model of myelodysplastic syndrome. The mutant zebrafish is an asxl1 mutant zebrafish, and the asxl1 mutant zebrafish is a deletion of 7 bases in the 12th exon of the zebrafish asxl1 gene from position 817 to position 823 or from position 798 to position 819 Zebrafish with a deletion of 22 bases. The present invention constructs for the first time a truncated asxl1 mutant similar to that of patients with blood myeloid tumors. After the mutation, it mainly affects the development of neutrophils in zebrafish, and has similar symptoms to myelodysplastic syndrome of human granulocytic dysplasia, and The present invention found for the first time that treatment with demethylase inhibitors against H3K4me3 can restore the phenotype of mutants. Therefore, this model may be used to study the pathogenesis of myelodysplastic syndromes caused by ASXL1 mutations and to screen drugs. new carrier.

Description

technical field [0001] The invention belongs to the field of biotechnology, and particularly relates to the application of a mutant zebrafish in preparing an animal model of myelodysplastic syndrome. Background technique [0002] Myeloid malignancies are clonal hematopoietic disorders caused by excessive proliferation, self-renewal or differentiation of hematopoietic stem cells (hematopoietic stem cells, HSC) or myeloid progenitor cells. In recent years, ASXL1 (additional sex combs-like 1) mutations have been widely found in many patients with hematological myeloid tumors, and the mutations are mainly concentrated on the 11th and 12th exons of the gene, that is, in the ASXL1 protein. Before the PHD functional domain, the mutation types are mainly frameshift mutations and nonsense mutations (①Carbuccia N, Murati A, TrouplinV, et al.Mutations of ASXL1gene in myeloproliferative neoplasms[J].Leukemia,2009,23(11):2183-2186 .② Gelsi-Boyer V, Trouplin V, Adelaide J, et al. Mutatio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/90C12N15/89A01K67/027
CPCA01K67/0275A01K2217/03A01K2227/40A01K2267/0331C12N15/902
Inventor 张文清方潇徐颂恩刘伟黄志斌
Owner SOUTH CHINA UNIV OF TECH
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