CRISPR ways to perform pooled screening

In this lecture, Dr. Andrew Bassett presented a comprehensive overview of pooled CRISPR screening as a high-throughput strategy for systematic interrogation of gene function at scale. He discussed why pooled screens have become central to functional genomics, including:

• the identification of essential genes,
• mapping disease-relevant genetic variation,
• discovery of therapeutic targets,
• and analysis of gene–gene interactions.

The lecture covered key experimental design principles, such as sgRNA library construction, choice of perturbation strategies (CRISPR knockout, CRISPRi, and CRISPRa), vector architectures, and requirements for library coverage and representation. Particular attention was given to model selection and delivery strategies across cancer cell lines, iPSC-derived systems, organoids, and in vivo models, as well as to maintaining experimental robustness during lentiviral transduction.

Dr. Bassett also reviewed available readout approaches, ranging from fitness-based assays and FACS to single-cell omics and image-based pooled screens, highlighting their respective strengths and limitations. Overall, the lecture emphasized best practices, common pitfalls, and the importance of follow-up validation, demonstrating how pooled CRISPR screening has already transformed functional genomics and continues to drive advances in disease research and precision medicine.