Fast Antibiotic resistance-Based gene editing of mammalian cells with CRISPR-Cas9 (FAB-CRISPR)

Petia Adarska; Eleanor Fox; Joshua Heyza; Carlo Barnaba; Jens Schmidt; Francesca Bottanelli

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arXiv · 2502.12675

Fast Antibiotic resistance-Based gene editing of mammalian cells with CRISPR-Cas9 (FAB-CRISPR)

Petia Adarska·Eleanor Fox·Joshua Heyza·Carlo Barnaba·Jens Schmidt·Francesca Bottanelli

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Abstract

Protein tagging with CRISPR-Cas9 enables the investigation of protein function in its native environment but is limited by low homology-directed repair (HDR) efficiency causing low knock-in rates. We present a detailed pipeline using HDR donor plasmids containing antibiotic resistance cassettes for rapid selection of gene-edited cells. Our protocol streamlines N- or C-terminal tagging in human cells, enabling HDR donor plasmid preparation in a single cloning step.

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Petia Adarska, Eleanor Fox, Joshua Heyza, Carlo Barnaba, Jens Schmidt, Francesca Bottanelli. 2025-02-18. Fast Antibiotic resistance-Based gene editing of mammalian cells with CRISPR-Cas9 (FAB-CRISPR). https://arxiv.org/abs/2502.12675

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