bioRxivCell Biology
A simple and efficient CRISPR technique for protein tagging
F. Zeng, V. Beck, S. Schuierer, I. Garnier, C. Manneville, C. Agarinis, L. Morelli, L. Quinn, J. Knehr, G. Roma, F. Bassilana, M. S. Nash
Abstract
Genetic knock-in using homology directed repair is an inefficient process, requiring 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% transfected cells with one single electroporation. The technique has proven effective in various cell types, 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 levels of a protein of interest by a luciferase assay in primary cells.
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