Fast Antibiotic resistance-Based gene editing of mammalian cells with CRISPR-Cas9 (FAB-CRISPR)
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arXiv
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| Autores principales: | , , , , , |
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| Formato: | Preprint |
| Publicado: |
2025
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| _version_ | 1866915158019801088 |
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| author | Adarska, Petia Fox, Eleanor Heyza, Joshua Barnaba, Carlo Schmidt, Jens Bottanelli, Francesca |
| author_facet | Adarska, Petia Fox, Eleanor Heyza, Joshua Barnaba, Carlo Schmidt, Jens Bottanelli, Francesca |
| contents | 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. |
| format | Preprint |
| id |
arxiv_https___arxiv_org_abs_2502_12675 |
| institution | arXiv |
| publishDate | 2025 |
| record_format | arxiv |
| spellingShingle | Fast Antibiotic resistance-Based gene editing of mammalian cells with CRISPR-Cas9 (FAB-CRISPR) Adarska, Petia Fox, Eleanor Heyza, Joshua Barnaba, Carlo Schmidt, Jens Bottanelli, Francesca Subcellular Processes 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. |
| title | Fast Antibiotic resistance-Based gene editing of mammalian cells with CRISPR-Cas9 (FAB-CRISPR) |
| topic | Subcellular Processes |
| url | https://arxiv.org/abs/2502.12675 |