Breeding and genotyping of T lymphocyte-conditional Spi1 knockout mice
10.19405/j.cnki.issn1000-1492.2024.04.006
- Author:
Huihui Wang
1
;
Xiangling Zhu
1
;
Xuming Wu
1
;
Huiru Zhang
1
;
Yuanyuan Zhou
1
;
Anqi Wang
1
;
Chong Liu
1
;
Jiajie Tu
1
Author Information
1. Institute of Clinical Pharmacology , Anhui Medical University , Key Laboratory of Anti-inflammatory and Immune Medicine , Ministry of Education , Collaborative Innovation Center of Anti-inflammatory and Immune Medicines , Hefei 230032
- Publication Type:Journal Article
- 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 Spi1 flox/flox mice to obtain Lck-Cre ×Spi1 flox/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 ×Spi1 flox/flox mouse genotype was stably inherited . Compared with Spi1 flox/flox mice , the expression level of PU. 1 was significantly reduced in splenic T cells of Lck-Cre ×Spi1 flox/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 .
- Full text:2024070222012467841T细胞条件性敲除Spi1基因小鼠的繁育及鉴定_王卉卉.pdf
