Preparation method and application of transgenic mouse with severe immunodeficiency

Abstract

The invention provides a preparation method and application of a transgenic mouse with severe immunodeficiency. The method comprises the step of knocking out a gene for encoding a gamma chain of an interleukin-2 receptor in a mouse chromosome through a CRISPR / CAS9 technology, exons 1 and 2 of a gene of a gamma chain of an interleukin-2 receptor are selected as knockout sites, and Cas9 messenger RNA and guide RNA are jointly injected into fertilized eggs of a mouse through microinjection. The mouse with severe immunodeficiency cannot generate T, B and NK immune cells, so that the greatest convenient condition is created for transplantation of human hematopoietic stem cells.

Description

technical field [0001] The invention relates to a preparation method and application of a severe immunodeficiency transgenic mouse, belonging to the field of biotechnology. Background technique [0002] The human immune response needs to be carried out in vivo, and due to ethical restrictions, it is not possible to directly use humans as experimental subjects. Rodents are small animal models widely used in experimental research. At present, most studies on the development, differentiation and function of the immune system come from these models. However, due to the existence of species specificity, the conclusions obtained from rodent models cannot be directly derived from humans. immune system. Human hematopoietic immune function research, infectious diseases, autoimmune diseases and tumor research are difficult to achieve dynamic observation in vivo. Some researchers use primates as substitutes, trying to make up for the huge species difference between humans and rodents...

AI Technical Summary

Problems solved by technology

However, there are many difficulties or risks in obtaining mice with NGF gene knock-in by conventional gene knock-in technology. Screening, a negative screening to obtain positive clones

After the homologous recombination vector enters the ES cells, it can randomly insert any sequence in the mouse genome, resulting in false positive results, resulting in the incorrect deletion of the mouse endogenous NGF gene

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