We have developed an in vivo screening platform using CRISPR/Cas9 to identify novel targets to improve anti-tumor therapies. In our pre-clinical model, genetic engineering of mouse t-cells prior to adoptive transfer into tumor-bearing mice allows us to identify genes that may be advantageous or deleterious to t-cell trafficking, persistence, and activation. Our sequencing analysis, combined with flow cytometry, creates a robust framework to analyze fitness of cells in a quantifiable manner. By using large genetic pools of potential targets, we can screen hundreds of targets in a single experiment.

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