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

Amy J. Heidersbach (), Kristel M. Dorighi, Javier A. Gomez, Ashley M. Jacobi and Benjamin Haley ()
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Amy J. Heidersbach: Genentech Inc.
Kristel M. Dorighi: Genentech Inc.
Javier A. Gomez: Integrated DNA Technology Inc
Ashley M. Jacobi: Integrated DNA Technology Inc
Benjamin Haley: Genentech Inc.

Nature Communications, 2023, vol. 14, issue 1, 1-10

Abstract: 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.

Date: 2023
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DOI: 10.1038/s41467-023-36452-w

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