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A simple and efficient CRISPR technique for protein tagging

Fanning Zeng, Valerie Beck, Sven Schuierer, Isabelle Garnier, Carole Manneville, Claudia Agarinis, Lapo Morelli, Lisa Quinn, Judith Knehr, Guglielmo Roma, Frederic Bassilana, Mark Nash
doi: https://doi.org/10.1101/597971
Fanning Zeng
Novartis Institutes for Biomedical Research, Basel, Switzerland
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  • For correspondence: fanning.zeng@novartis.com
Valerie Beck
Novartis Institutes for Biomedical Research, Basel, Switzerland
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Sven Schuierer
Novartis Institutes for Biomedical Research, Basel, Switzerland
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Isabelle Garnier
Novartis Institutes for Biomedical Research, Basel, Switzerland
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Carole Manneville
Novartis Institutes for Biomedical Research, Basel, Switzerland
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Claudia Agarinis
Novartis Institutes for Biomedical Research, Basel, Switzerland
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Lapo Morelli
Novartis Institutes for Biomedical Research, Basel, Switzerland
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Lisa Quinn
Novartis Institutes for Biomedical Research, Basel, Switzerland
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Judith Knehr
Novartis Institutes for Biomedical Research, Basel, Switzerland
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Guglielmo Roma
Novartis Institutes for Biomedical Research, Basel, Switzerland
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Frederic Bassilana
Novartis Institutes for Biomedical Research, Basel, Switzerland
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Mark Nash
Novartis Institutes for Biomedical Research, Basel, Switzerland
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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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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted April 05, 2019.
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A simple and efficient CRISPR technique for protein tagging
Fanning Zeng, Valerie Beck, Sven Schuierer, Isabelle Garnier, Carole Manneville, Claudia Agarinis, Lapo Morelli, Lisa Quinn, Judith Knehr, Guglielmo Roma, Frederic Bassilana, Mark Nash
bioRxiv 597971; doi: https://doi.org/10.1101/597971
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A simple and efficient CRISPR technique for protein tagging
Fanning Zeng, Valerie Beck, Sven Schuierer, Isabelle Garnier, Carole Manneville, Claudia Agarinis, Lapo Morelli, Lisa Quinn, Judith Knehr, Guglielmo Roma, Frederic Bassilana, Mark Nash
bioRxiv 597971; doi: https://doi.org/10.1101/597971

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