Related Experiment Video
Updated: Sep 15, 2025
Virus Delivery of CRISPR Guides to the Murine Prostate for Gene Alteration
Published on: April 27, 2018
The Establishment of Prostate-specific, SKP2 Humanized Mice by CRISPR Knock-in Method Reveals Neoplastic Initiation
Abstract:
A recent study has shown that SKP2 inactivation can prevent cancer initiation by extension of total cell cycle duration without perturbing normal division, which suggests a new strategy for cancer prevention. However, direct in vivo evidence for human SKP2 on cancer initiation and prostatic microenvironment is still lacking and a prostate-specific SKP2 humanized mouse model is critical for developing prostate cancer immunoprevention approaches through targeting human SKP2. We therefore have established a prostate-specific human SKP2 (h SKP2 ) knock-in mouse model by a CRISPR knock-in approach. Overexpression of h SKP2, which is driven by an endogenous mouse probasin promoter, induces prostatic lesions including hyperplasia, mouse prostate intraepithelial neoplasia (mPIN), and low-grade carcinoma and increases prostate weights. Transcriptional profiling by RNA-sequencing analysis revealed significant gene expression alterations in epithelial to mesenchymal transition (EMT), extracellular matrix, and interferon signaling in the prostate of h SKP2 knock-in mice compared to wild-type mice. Single cell deconvolution showed an increase of fibroblasts population and a decrease of CD8 + T cell and B cell populations in the prostate of hSKP2 -knock-in mice. Consistently with these results from the SKP2 humanized mouse, overexpression of hSKP2 in human prostate cancer PC3 cells markedly increased cell migration and invasion and induced the gene expression of EMT and interferon pathways, including FMOD, THY1, PFKP, USP18, IL15, etc. In addition, paired prostate organoids were derived from SKP2 humanized and wild-type mice for drug screening and validated by known SKP2 inhibitors, Flavokawain A and C1. Both of which selectively decrease the viability and alter the morphologies of organoids of h SKP2 knock-in rather than wild-type mice. Our studies provide a well-characterized prostate-specific h SKP2 knock-in mouse model and offer new mechanistic insights for understanding the oncogenic role of SKP2 in shaping the prostatic microenvironment during early carcinogenesis.
Insights
A new prostate-specific human SKP2 mouse model reveals SKP2’s role in prostate cancer initiation and microenvironment changes. This model aids in developing targeted cancer prevention strategies and drug screening for prostate cancer.
Area of Science:
- Oncology
- Genetics
- Molecular Biology
Background:
- SKP2 (S-phase kinase-associated protein 2) inactivation may prevent cancer by extending cell cycle duration.
- Direct in vivo evidence for human SKP2's role in prostate cancer initiation and its microenvironment is lacking.
- A prostate-specific SKP2 humanized mouse model is crucial for developing targeted cancer prevention strategies.
Purpose of the Study:
- To establish a prostate-specific human SKP2 (hSKP2) knock-in mouse model.
- To investigate the oncogenic role of hSKP2 in prostate carcinogenesis and its impact on the prostatic microenvironment.
- To provide a platform for drug screening against SKP2 in prostate cancer.
Main Methods:
- CRISPR-Cas9 gene editing to create a prostate-specific hSKP2 knock-in mouse model.
- RNA-sequencing for transcriptional profiling of prostate tissues.
- Single-cell deconvolution to analyze cell populations.
- In vitro studies using human prostate cancer cells and mouse organoids.
- Drug screening using SKP2 inhibitors.
Main Results:
- Overexpression of hSKP2 induced prostatic lesions (hyperplasia, mPIN, carcinoma) and increased prostate weight.
- Transcriptional profiling revealed alterations in epithelial to mesenchymal transition (EMT), extracellular matrix, and interferon signaling.
- Single-cell analysis showed increased fibroblasts and decreased CD8+ T and B cells in hSKP2 knock-in mice.
- hSKP2 overexpression increased human prostate cancer cell migration, invasion, and EMT gene expression.
- SKP2 inhibitors selectively reduced the viability of hSKP2 knock-in mouse organoids.
Conclusions:
- The established prostate-specific hSKP2 knock-in mouse model effectively recapitulates key aspects of prostate carcinogenesis.
- hSKP2 plays a significant role in shaping the prostatic microenvironment during early carcinogenesis.
- This model serves as a valuable tool for understanding SKP2's oncogenic functions and for preclinical drug development in prostate cancer.
