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A Simple and Efficient CRISPR Technique for Protein Tagging.

ABSTRACT: 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.

SUBMITTER: Zeng F

PROVIDER: S-EPMC7762194 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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A Simple and Efficient CRISPR Technique for Protein Tagging.

Zeng Fanning F Beck Valerie V Schuierer Sven S Garnier Isabelle I Manneville Carole C Agarinis Claudia C Morelli Lapo L Quinn Lisa L Knehr Judith J Roma Guglielmo G Bassilana Frederic F Nash Mark M

Cells 20201205 12

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 lo ...[more]

PMID: 33291479

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