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
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author Zeng, Fanning
Beck, Valerie
Schuierer, Sven
Garnier, Isabelle
Manneville, Carole
Agarinis, Claudia
Morelli, Lapo
Quinn, Lisa
Knehr, Judith
Roma, Guglielmo
Bassilana, Frederic
Nash, Mark
author_facet Zeng, Fanning
Beck, Valerie
Schuierer, Sven
Garnier, Isabelle
Manneville, Carole
Agarinis, Claudia
Morelli, Lapo
Quinn, Lisa
Knehr, Judith
Roma, Guglielmo
Bassilana, Frederic
Nash, Mark
author_sort Zeng, Fanning
collection PubMed
description 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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spelling pubmed-77621942020-12-26 A Simple and Efficient CRISPR Technique for Protein Tagging Zeng, Fanning Beck, Valerie Schuierer, Sven Garnier, Isabelle Manneville, Carole Agarinis, Claudia Morelli, Lapo Quinn, Lisa Knehr, Judith Roma, Guglielmo Bassilana, Frederic Nash, Mark Cells Article 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. MDPI 2020-12-05 /pmc/articles/PMC7762194/ /pubmed/33291479 http://dx.doi.org/10.3390/cells9122618 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zeng, Fanning
Beck, Valerie
Schuierer, Sven
Garnier, Isabelle
Manneville, Carole
Agarinis, Claudia
Morelli, Lapo
Quinn, Lisa
Knehr, Judith
Roma, Guglielmo
Bassilana, Frederic
Nash, Mark
A Simple and Efficient CRISPR Technique for Protein Tagging
title A Simple and Efficient CRISPR Technique for Protein Tagging
title_full A Simple and Efficient CRISPR Technique for Protein Tagging
title_fullStr A Simple and Efficient CRISPR Technique for Protein Tagging
title_full_unstemmed A Simple and Efficient CRISPR Technique for Protein Tagging
title_short A Simple and Efficient CRISPR Technique for Protein Tagging
title_sort simple and efficient crispr technique for protein tagging
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7762194/
https://www.ncbi.nlm.nih.gov/pubmed/33291479
http://dx.doi.org/10.3390/cells9122618
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