Cultivation method for mouse with kidney podocyte specific knockout lncRNA DLX6-os1 transgene

Abstract

The invention relates to a cultivation method for a mouse with a kidney podocyte specific knockout lncRNA DLX6-os1 transgene. The method can effectively achieve the effect of studying the action mechanism of podocyte injuries in different kidney diseases; according to the technical scheme, multi-generation crossbreeding is performed on an FLOX-DLX6-os1 transgenic mouse constructed in the earlier stage and another transgenic mouse carrying a podocyte specific NPHS2-Cre gene through a genetic engineering technology, and the cultivation of the mouse with the kidney podocyte specific knockout lncRNA DLX6-os1 transgene is achieved. The cultivation method for the mouse with the kidney podocyte specific knockout lncRNA DLX6-os1 transgene provides a very good animal model for defining the role ofthe lncRNA DLX6-os1 in the podocyte injuries, effectively achieves the effect of studying the action mechanism of the podocyte injuries in the different kidney diseases, and is an innovation of the cultivation method of transgenic mice.

Description

technical field [0001] The invention relates to medical biology, in particular to a breeding method for kidney podocyte-specific knockout lncRNA DLX6-os1 transgenic mice. Background technique [0002] The filtration function of the kidney is crucial to maintaining the normal operation of the body, and the maintenance of its filtration function requires a filtration barrier to function. One of the most important components of the filtration barrier is the podocyte. The health of podocytes is related to the normal maintenance of kidney function and even the whole body function. In the disease state, if podocytes are damaged, podocyte hypertrophy, apoptosis, or foot process fusion will lead to a decline in the filtration function of podocytes, the most direct clinical manifestation is proteinuria in patients, and the appearance of proteinuria will It will aggravate the damage of the entire kidney. It has been found that one of the keys to the occurrence and development of var...

AI Technical Summary

Problems solved by technology

[0003] Previous research has found that the expression of LncRNA DLX6-AS1 (DLX6 antisense RNA 1: RefSeq GeneID 285987, corresponding to mouse Dlx6-os1) is increased in renal tissues of patients with diabetic nephropathy and IgA nephropathy, especially in podocytes The expression of DLX6-os1 was higher than that of the normal control group. It was further verified by animal models of kidney disease and cell models. It was found that the expression level of DLX6-os1 was positively correlated with the increase of urinary protein in mice, and positively correlated with the degree of podocyte damage. Further experiments found that in Overexpression of lncRNA DLX6-os1 in normal podocytes can lead to podocyte injury and proteinuria in mice, indicating that lncRNA DLX6-os1 is closely related to kidney injury and podocyte injury, but the specific mechanism and role are unknown

[0004] In order to further clarify the important role of this lncRNA in podocytes, it is necessary to change the expression level of DLX6-os1 in podocytes of experimental animals. At present, the commonly used method in the world is to inject viruses carrying lncRNA into corresponding tissues and organs in an attempt to change the expression of this lncRNA Even if the lentiviral vector specifically expressed in podocytes is used, there are still many problems: 1. The infection efficiency of the virus is low, and the effect of effectively changing the expression level of lncRNA in specific cells cannot be achieved; 2. Some lncRNA sequences are longer , not suitable for packaging virus or causing low titer of virus, not suitable for infecting animals; 3. The effect in specific target cells is poor

[0005] Therefore, the previously established transgenic mice carrying FLOX-DLX6-os1 were further genetically modified by genetic engineering technology, and a method of breeding kidney podocyte-specific knockout lncRNA DLX6-os1 transgenic mice (flox / flox, NPHS2-Cre) was invented. , providing a very good animal model for clarifying the role of lncRNA DLX6-os1 in podocyte injury, and effectively solving the problem of studying the mechanism of podocyte injury in different kidney diseases, but there has been no public report so far

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