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 |
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Format: | Artikel |
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. |
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ISSN: | 2073-4409 2073-4409 |
DOI: | 10.3390/cells9122618 |