Construction, breeding, and gene identification of micro RNA - 22 - 3p knockout mice
10.19405/j.cnki.issn1000-1492.2025.06.011
- Author:
Anqi Wang
1
;
Huiru Zhang
1
;
Yuanyuan Zhou
1
;
Chong Liu
1
;
Yizhao Chen
1
;
Jiajie Tu
1
Author Information
1. College of Pharmaceutical Sciences , Anhui Medical University , Hefei 230032
- Publication Type:Journal Article
- Keywords:
miR-22;
CRISPR/Cas 9;
gene knockout;
polymerase chain reaction;
agarose gel electrophoresis;
genotype identification;
NLRP3
- From:
Acta Universitatis Medicinalis Anhui
2025;60(6):1052-1058
- CountryChina
- Language:Chinese
-
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 generation 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 performed 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 expression 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.
- Full text:2026030315334719783微小RNA-22-3p基因敲除小鼠的构建、繁育和基因鉴定_王安琪.pdf
