安徽医科大学学报2025，Vol.60Issue(6)：1015-1021,7.DOI:10.19405/j.cnki.issn1000-1492.2025.06.006
IL-9R CDS基因人源化小鼠模型构建与验证
Construction and verification of humanized mouse model of IL-9R CDS gene
摘要
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 ge-nome 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 suc-cessfully constructed and characterized.关键词
基因人源化/白细胞介素9受体/CRISPR/Cas9/聚合酶链式反应/免疫印迹Key words
gene humanization/IL-9R/CRISPR/Cas9/PCR/Western blot分类
临床医学引用本文复制引用
刘崇,周园园,薛慧,薛梓萌,陈维乐,涂佳杰..IL-9R CDS基因人源化小鼠模型构建与验证[J].安徽医科大学学报,2025,60(6):1015-1021,7.基金项目
安徽省高校杰出青年科研项目(编号:2022AH020052) Distinguished Young Scholars Support Project of Anhui Educational Committee(No.2022AH020052) （编号:2022AH020052）
