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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Detalles Bibliográficos
Autores principales: Zeng, Fanning, Beck, Valerie, Schuierer, Sven, Garnier, Isabelle, Manneville, Carole, Agarinis, Claudia, Morelli, Lapo, Quinn, Lisa, Knehr, Judith, Roma, Guglielmo, Bassilana, Frederic, Nash, Mark
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7762194/
https://www.ncbi.nlm.nih.gov/pubmed/33291479
http://dx.doi.org/10.3390/cells9122618
Descripción
Sumario: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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