In vivo CRISPR screening protocol to identify metastasis mediators using iteratively selected mouse models

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Xi Wang

doi:10.1016/j.xpro.2025.104042

In vivo CRISPR screening protocol to identify metastasis mediators using iteratively selected mouse models

Xi Wang · 2025-09-17

In vivo CRISPR screens uncover metastasis genes in native contexts, surpassing in vitro model limitations. Here, we present a protocol to identify metastasis-driving genes in ovarian cancer using an in vivo CRISPR screening technique. Key steps include single-guide RNA (sgRNA) library design and validation, lentiviral transduction, establishment of metastatic mouse models, tissue collection, sgRNA amplification for sequencing, bioinformatics-based candidate gene identification, and functional validation. For complete details on the use and execution of this protocol, please refer to Wang et al.

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In vivo CRISPR screens uncover metastasis genes in native contexts, surpassing in vitro model limitations, and identifying metastasis-driving genes in ovarian cancer using an in vivo CRISPR screening technique.

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In vivo CRISPR screening protocol to identify metastasis mediators using iteratively selected mouse models

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