Construction and verification of humanized mouse model of IL - 9R CDS gene
10.19405/j.cnki.issn1000-1492.2025.06.006
- Author:
Chong Liu
1
;
Yuanyuan Zhou
1
;
Hui Xue
1
;
Zimeng Xue
1
;
Weile Chen
1
;
Jiajie Tu
1
Author Information
1. College of Pharmaceutical Sciences , Anhui Medical University , Hefei 230032
- Publication Type:Journal Article
- Keywords:
gene humanization;
IL-9R;
CRISPR/Cas9;
PCR;
Western blot
- From:
Acta Universitatis Medicinalis Anhui
2025;60(6):1015-1021
- CountryChina
- Language:Chinese
-
Abstract:
Objective :To construct a humanized mouse model of the interleukin-9 receptor(IL-9R) coding DNA sequence(CDS) gene and to verify the genotype and IL-9R expression in mice.
Methods : The CRISPR/Cas9 genome editing technology was used to replace the exon 2-7 fragment of the il-9r gene in mouse embryonic stem cells with the corresponding human IL-9R sequence. After verifying the completion of the gene fragment replacement, tetraploid embryos were constructed and microinjected back into the oviducts of surrogate mice. Through surrogacy by female mice, homozygous humanized mice were obtained. DNA was extracted from the homozygous humanized mice IL-9R CDS gene, and their genotypes were identified by agarose gel electrophoresis after PCR amplification. Western blot was used to detect the expression of IL-9R in the spleen and thymus of homozygous humanized mice with either wild-type(WT) or IL-9R gene humanization.
Results :Gel electrophoresis after PCR amplification showed that mice with only a 1 805 bp band amplified using WT primers were wild-type, while mice with 2 553 bp and 2 340 bp bands amplified using 5KI and 3KI primers, respectively, were homozygous humanized mice with IL-9R CDS gene. Western blot results indicated that the tissues of homozygous humanized mice model with IL-9R CDS gene expressed IL-9R significantly.
Conclusion:The humanized mouse model with IL-9R CDS gene has been successfully constructed and characterized.
- Full text:2026030216032203484IL-9RCDS基因人源化小鼠模型构建与验证_刘崇.pdf
