A versatile, high-efficiency platform for CRISPR-based gene activation

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Abstract

CRISPR-mediated transcriptional activation (CRISPRa) is a powerful technology for inducing gene expression from endogenous loci with exciting applications in high throughput gain-of-function genomic screens and the engineering of cell-based models. However, current strategies for generating potent, stable, CRISPRa-competent cell lines present limitations for the broad utility of this approach. Here, we provide a high-efficiency, self-selecting CRISPRa enrichment strategy, which combined with piggyBac transposon technology enables rapid production of CRISPRa-ready cell populations compatible with a variety of downstream assays. We complement this with an optimized guide RNA scaffold that significantly enhances CRISPRa functionality. Finally, we describe a synthetic guide RNA tool set that enables transient, population-wide gene activation when used with the self-selecting CRISPRa system. Taken together, this versatile platform greatly enhances the potential for CRISPRa across a wide variety of cellular contexts.

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Heidersbach, A. J., Dorighi, K. M., Gomez, J. A., Jacobi, A. M., & Haley, B. (2023). A versatile, high-efficiency platform for CRISPR-based gene activation. Nature Communications, 14(1). https://doi.org/10.1038/s41467-023-36452-w

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