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Constructing a mouse model against MLL leukemia by changing the expression level of hoxb13 and the method and application of the model construction

A hoxb13, construction method technology, applied in the directions of application, chemical instruments and methods, botanical equipment and methods, etc., can solve the problems of luminal epithelial cell polarity and secretory function loss, unresearched, and ventral prostate lobe morphological defects. , to achieve good application prospects and easy-to-obtain results

Active Publication Date: 2022-02-11
INST OF HEMATOLOGY & BLOOD DISEASES HOSPITAL CHINESE ACADEMY OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
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Problems solved by technology

In 2003, mice homozygous for the Hoxb13 mutation were found to have defects in ventral prostatic lobe morphology, as well as loss of luminal epithelial cell polarity and secretory function
In MLL-rearranged leukemia, the change of Hoxb13 expression level has no effect on the regulation of tumor growth and development.

Method used

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  • Constructing a mouse model against MLL leukemia by changing the expression level of hoxb13 and the method and application of the model construction
  • Constructing a mouse model against MLL leukemia by changing the expression level of hoxb13 and the method and application of the model construction
  • Constructing a mouse model against MLL leukemia by changing the expression level of hoxb13 and the method and application of the model construction

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Embodiment Construction

[0034] The present invention will be described in detail below in conjunction with the accompanying drawings and embodiments.

[0035] 1. Construction of MLL-AF9 leukemia mice

[0036] 1.1 Enrichment of mouse bone marrow c-Kit+ cells

[0037] 1) C57 mice were sacrificed by vertebral dislocation, soaked and disinfected with 75% ethanol solution, and surgical instruments and sterile gauze blocks after autoclaving were used to dissect the bilateral hindlimb bones of the mice, and the iliac bones were isolated sequentially. bone, femur, and tibia, and use gauze blocks to peel off the muscle on the outer surface of the bone as much as possible, so as to prevent the muscle from affecting the subsequent flushing of the bone marrow cavity. Use a 1ml BD syringe in 3ml PBE liquid to repeatedly flush the bone marrow cavity from both ends of the bone to obtain bone marrow cells.

[0038] 2) Filter 3ml of the bone marrow cell flushing solution collected through a nylon membrane with a po...

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Abstract

The invention discloses a mouse model for resisting MLL leukemia by changing the expression level of Hoxb13, a method for constructing the model and an application thereof. The present invention utilizes lentivirus-mediated gene introduction technology to introduce the shRNA plasmid specifically knocking down the mRNA produced by Hoxb13 gene transcription into MLL leukemia cells, and then transplant it to recipient mice irradiated with a sublethal dose to obtain resistance to MLL A mouse model of leukemia. The mice after disease remission showed a significantly prolonged survival period, and decreased proliferation and colony formation abilities of leukemia cells. The mouse model of the invention can easily, quickly and efficiently realize the qualitative and / or quantitative evaluation of the potential medicine for MLL leukemia, and has broad application prospects.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a mouse model for resisting MLL leukemia by changing the expression level of Hoxb13, a model construction method and application. Background technique [0002] Leukemia is a malignant tumor of the hematopoietic system and one of the top ten high-incidence malignant tumors in my country. Leukemic cells originate from hematopoietic stem and progenitor cells, which can lead to the accumulation of leukemia cells in the blood system and invade other tissues and organs through malignant proliferation. The clinical manifestations are anemia, bleeding, infection, fever, and hepatosplenomegaly. It can lead to the death of patients and seriously endanger human health and life safety. There are many hypotheses about the formation of leukemia clones. The current mainstream view is that a single hematopoietic stem and progenitor cell undergoes malignant transformation to produce leukemia stem ce...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/867C12N5/10A01K67/027
CPCC12N15/113C12N15/86C07K14/47A01K67/0271C12N2310/14C12N2740/15043A01K2207/12A01K2227/105A01K2267/0331
Inventor 袁卫平初雅婧施均陈仰鹏汪晓敏
Owner INST OF HEMATOLOGY & BLOOD DISEASES HOSPITAL CHINESE ACADEMY OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
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