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Production of Genetically Engineered Golden Syrian Hamsters by Pronuclear Injection of the CRISPR/Cas9 Complex
Published on: January 9, 2018
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Characterization of SHARPIN knockout Syrian hamsters developed using CRISPR/Cas9 system.
Jinxin Miao1, Tianfeng Lan2, Haoran Guo2
1Academy of Chinese Medicine Science, Henan University of Chinese Medicine, Zhengzhou, Henan, People's Republic of China.
Animal Models and Experimental Medicine
|September 13, 2022
Summary
Researchers created a novel SHARPIN knockout hamster model using CRISPR/Cas9 gene editing. This model exhibits spleen enlargement and eosinophil infiltration, offering new insights into SHARPIN
Area of Science:
- Genetics and Genomics
- Immunology
- Molecular Biology
Background:
- SHARPIN (SHANK-associated RH domain interactor) is a key regulator of the NF-κB signaling pathway within the linear ubiquitination complex.
- Understanding SHARPIN's precise function is crucial for dissecting inflammatory and immune responses.
Purpose of the Study:
- To establish a novel, genetically engineered Syrian hamster model with targeted SHARPIN gene disruption.
- To investigate the physiological and molecular consequences of SHARPIN deficiency in vivo.
Main Methods:
- CRISPR/Cas9 gene editing was employed to create single-guide RNA targeting the SHARPIN gene.
- Genotyping, Western blotting, organ weight analysis, histopathology, and gene expression analysis (RT-qPCR) were performed on mutant and wild-type hamsters.
Main Results:
- Germline-transmitted SHARPIN mutations were confirmed, leading to undetectable SHARPIN protein in knockout (KO) hamsters.
- SHARPIN-/- hamsters displayed spleen enlargement, thymic and lymph node reduction, and significant eosinophil infiltration across multiple organs.
- Downregulation of CD94/CD22 and upregulation of CCR3/CCL11/Il4/Il13 were observed, alongside diminished NF-κB signaling.
Conclusions:
- A novel SHARPIN knockout hamster model was successfully generated using CRISPR/Cas9 technology.
- The observed abnormalities in lymphoid organ development and widespread eosinophil infiltration highlight SHARPIN's role in regulating inflammation and immune homeostasis.
