Mouse pluripotent stem cell line for erythroid specific high-expression GFP (Green Fluorescent Protein) and construction method of mouse pluripotent stem cell line

Abstract

The invention discloses a mouse pluripotent stem cell line for an erythroid specific high-expression GFP (Green Fluorescent Protein) and a construction method of the mouse pluripotent stem cell line.The mouse pluripotent stem cell line is prepared by the following steps: injecting an erythroid high-expression GFP vector into male pronucleus of Kunming white mouse fertilized eggs in a micro-injection manner, cultivating transgenic mice, verifying expressions of inserted genes at specific tissue cells, reproducing the transgenic mice, stabilizing the copy number of exogenous genes of generationmice of the transgenic mice, and preparing the mouse pluripotent stem cell line by blastaea. The mouse pluripotent stem cell line disclosed by the invention has important cytological value for researching in-vivo and in-vitro formation and evolution of blood cells, which contributes to uncovering the hematopoietic stem formation and complicated influence effect of the hematopoietic microenvironment and pushing the research process of differentiating pluripotent stem cells to transplantable hematopoietic stem cells in vitro.

Description

technical field [0001] The invention belongs to the technical field of induced stem cells, and in particular relates to a mouse pluripotent stem cell line with erythroid-specific high expression of GFP and a construction method thereof. Background technique [0002] The first mouse transgenic embryonic stem cells (ESCs) were obtained in 1981. ESCs have self-renewal and multi-directional differentiation potential. It has been confirmed that ESCs can differentiate into all tissue cells by using tetraploid compensation technology. In terms of in vitro differentiation, researchers have successfully induced ESCs to form pancreatic cells, nerve cells, cardiomyocytes, and hematopoietic cells. Stem cells and other tissue cells. These characteristics of ESCs have promoted the research of functional genes. ESCs carrying specific exogenous genes can study the basic functions and regulatory mechanisms of genes in vivo and in vitro, and can develop molecular biological farms to explore...

AI Technical Summary

Problems solved by technology

However, the commonly used transgenic technology cannot fully achieve directional integration, and the copy number of the integrated gene cannot be effectively controlled, which leads to differences in the expression of exogenous genes among transgenic mice and among the offspring of transgenic mice

[0004] Conventional transgenic embryonic stem cells directly introduce exogenous genes into ESCs. This method often results in random insertion of exogenous genes, inconsistencies in the copy number of transgenes in each cell clone, and expression silencing of inserted genes.

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