Breeding and genotyping of T lymphocyte-conditional Spi1 knockout mice
10.19405/j.cnki.issn1000-1492.2024.04.006
- VernacularTitle:T细胞条件性敲除Spi1基因小鼠的繁育及鉴定
- Author:
Huihui WANG
1
;
Xiangling ZHU
;
Xuming WU
;
Huiru ZHANG
;
Yuanyuan ZHOU
;
Anqi WANG
;
Chong LIU
;
Jiajie TU
Author Information
1. 安徽医科大学临床药理研究所,抗炎免疫药物教育部重点实验室,抗炎免疫药物安徽协同创新中心,合肥 230032
- Keywords:
Spi1;
Cre/LoxP system;
CRISPR/Cas9 technology;
T cells;
PU.1;
conditional knockout
- From:
Acta Universitatis Medicinalis Anhui
2024;59(4):595-599
- CountryChina
- Language:Chinese
-
Abstract:
Objective To breed and identify the T lymphocyte-conditional Spi1 knockout mice for the further in-vestgation of the specific role of Spi1-encoded protein PU.1.Methods The Lck-Cre mice were mated with Spi1flox/flox mice to obtain Lck-Cre×Spi1flox/flox mice(T lymphocyte-specific Spi1 knockout mice),and the genotype was determined by polymerase chain reaction(PCR)and agarose gel electrophoresis.Magnetic beads were used to sort out the splenic T lymphocytes,and the knockdown efficiency of PU.1 in T cells was detected by Western blot,quantitative real-time PCR(qPCR)and flow cytometry.Results The Lck-Cre×Spi1flox/flox mouse genotype was stably inherited.Compared with Spi1flox/flox mice,the expression level of PU.1 was significantly reduced in splenic T cells of Lck-Cre×Spi1flox/flox mice.Conclusion In this study,the T lymphocyte-specific Spi1 knockout mice was successfully constructed by applying Cre/LoxP system and CRISPR/Cas9 technology,which provided a reliable an-imal model for the subsequent experiments of the specific role of PU.1 in T cell-related diseases.
