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

2025/02/18 by Petia Adarska, Adarska, Petia, Eleanor Fox +10 · 2 voices
Biochemistry, Genetics and Molecular Biology · #CRISPR and Genetic Engineering

paper · pdf · doi:10.48550/arxiv.2502.12675

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.

Citations

• A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity

Discussions

• Long in the making but our protocol for gene editing is online! FAB CRISPR is super easy and allows the generation of N and C terminal fusions with TurboID, SNAP, Halo and mStayGold. A single cloninf [bsky, 69 points, 3 comments]
• Petia Adarska, Eleanor Fox, Joshua Heyza, Carlo Barnaba, Jens Schmidt, Francesca Bottanelli: Fast Antibiotic resistance-Based gene editing of mammalian cells with CRISPR-Cas9 (FAB-CRISPR) https://arxi [bsky, 0 points, 0 comments]

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