A Simple and Efficient CRISPR Technique for Protein Tagging

Genetic knock-in using homology-directed repair is an inefficient process, requiring the selection of few modified cells and hindering its application to primary cells. Here, we describe Homology independent gene Tagging (HiTag), a method to tag a protein of interest by CRISPR in up to 66% of transf...

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Veröffentlicht in:Cells (Basel, Switzerland) Switzerland), 2020-12, Vol.9 (12), p.2618, Article 2618
Hauptverfasser: Zeng, Fanning, Beck, Valerie, Schuierer, Sven, Garnier, Isabelle, Manneville, Carole, Agarinis, Claudia, Morelli, Lapo, Quinn, Lisa, Knehr, Judith, Roma, Guglielmo, Bassilana, Frederic, Nash, Mark
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Sprache:eng
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Zusammenfassung:Genetic knock-in using homology-directed repair is an inefficient process, requiring the selection of few modified cells and hindering its application to primary cells. Here, we describe Homology independent gene Tagging (HiTag), a method to tag a protein of interest by CRISPR in up to 66% of transfected cells with one single electroporation. The technique has proven effective in various cell types and can be used to knock in a fluorescent protein for live cell imaging, to modify the cellular location of a target protein and to monitor the levels of a protein of interest by a luciferase assay in primary cells.
ISSN:2073-4409
2073-4409
DOI:10.3390/cells9122618