安徽医科大学学报2025，Vol.60Issue(6)：1052-1058,7.DOI:10.19405/j.cnki.issn1000-1492.2025.06.011
微小RNA-22-3p基因敲除小鼠的构建、繁育和基因鉴定
Construction,breeding,and gene identification of microRNA-22-3p knockout mice
摘要
Abstract
Objective To construct microRNA(miR)-22 gene knockout(miR-22-/-)mice using CRISPR/Cas 9 technology,to breed miR-22-/-mice and to identify their genotypes.Methods In this experiment,CRISPR/Cas 9 technology was used to construct miR-22-/-genetically engineered mice.After gene identification,the F0 gener-ation miR-22-/-mice were mated with wild-type mice in the same litter to obtain F1 generation miR-22-/-mice.The miR-22 knockout efficiency was analyzed at the RNA level by real-time fluorescence quantitative polymerase chain reaction(qPCR).Western blot was used to detect the interaction between miR-22 and target genes.Results miR-22-/-mice were successfully constructed using CRISPR/Cas 9 technology,gene identification was per-formed on the bred mice,and three stable genotypes of miR-22+/+,miR-22+/-,and miR-22-/-were identified.The real-time fluorescence quantitative PCR detection results confirmed that miR-22-/-mice showed almost no ex-pression of miR-22 in the heart,liver,lung,kidney,spleen,and thymus tissues compared to wild-type mice in the same litter.Western blot analysis showed that the relative expression level of NLRP3 protein in miR-22-/-mouse tissues was lower than that in wild-type mice.Conclusion A miR-22-/-mouse model is successfully constructed,and stable genetic homozygous miR-22-/-mice is obtained.This indicates that miR-22 has an inhibitory effect on the downstream target gene NLRP3.关键词
miR-22/CRISPR/Cas 9/基因敲除/聚合酶链式反应/琼脂糖凝胶电泳/基因型鉴定/NLRP3Key words
miR-22/CRISPR/Cas 9/gene knockout/polymerase chain reaction/agarose gel electrophoresis/genotype identification/NLRP3分类
基础医学引用本文复制引用
王安琪,张慧茹,周园园,刘崇,陈义昭,涂佳杰..微小RNA-22-3p基因敲除小鼠的构建、繁育和基因鉴定[J].安徽医科大学学报,2025,60(6):1052-1058,7.基金项目
安徽省高校杰出青年科研项目(编号:2022AH020052) Natural Science Research Project of Anhui Educational Committee(No.2022AH020052) （编号:2022AH020052）
