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

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Amy Heidersbach , Kristel M. Dorighi , Javier A. Gomez , Ashley M. Jacobi , Benjamin Haley ·

YOU? · · 2023 · Open Access · · DOI: https://doi.org/10.1038/s41467-023-36452-w

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.

Related Topics

Crispr

Cas9

Computational Biology

Genome Editing

Computer Science

Biology

Concepts

CRISPR Cas9 Computational biology Genome engineering Computer science Biology Gene Genetics

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Type: article
Language: en
Landing Page: https://doi.org/10.1038/s41467-023-36452-w
PDF: https://www.nature.com/articles/s41467-023-36452-w.pdf
OA Status: gold
Cited By: 52
References: 31
Related Works: 10
OpenAlex ID: https://openalex.org/W4321240427

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OpenAlex ID: https://openalex.org/W4321240427

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DOI: https://doi.org/10.1038/s41467-023-36452-w

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

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Type: article

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Language: en

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Publication year: 2023

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Publication date: 2023-02-17

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Authors: Amy Heidersbach, Kristel M. Dorighi, Javier A. Gomez, Ashley M. Jacobi, Benjamin Haley

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Landing page: https://doi.org/10.1038/s41467-023-36452-w

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PDF URL: https://www.nature.com/articles/s41467-023-36452-w.pdf

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Open access: Yes

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OA status: gold

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OA URL: https://www.nature.com/articles/s41467-023-36452-w.pdf

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Concepts: CRISPR, Cas9, Computational biology, Genome engineering, Computer science, Biology, Gene, Genetics

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Cited by: 52

Total citation count in OpenAlex
Citations by year (recent): 2025: 21, 2024: 24, 2023: 7

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References (count): 31

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Related works (count): 10

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