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Versatile labeling and detection of endogenous proteins using tag-assisted split enzyme complementation

View ORCID ProfileSuraj Makhija, View ORCID ProfileDavid Brown, View ORCID ProfileStruan Bourke, Yina Wang, View ORCID ProfileShuqin Zhou, View ORCID ProfileRachel Rudlaff, View ORCID ProfileRasmi Cheloor-Kovilakam, View ORCID ProfileBo Huang
doi: https://doi.org/10.1101/2020.12.01.407072
Suraj Makhija
1UC Berkeley - UCSF Joint Graduate Program in Bioengineering, University of California, San Francisco, San Francisco, CA 94143 USA
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David Brown
2Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94143 USA
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Struan Bourke
2Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94143 USA
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Yina Wang
2Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94143 USA
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Shuqin Zhou
3Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, CA 94143 USA
4School of Pharmacy, Tsinghua University, Beijing 100872 China
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Rachel Rudlaff
2Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94143 USA
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Rasmi Cheloor-Kovilakam
2Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94143 USA
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Bo Huang
2Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94143 USA
5Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA 94143 USA
6Chan Zuckerberg Biohub, San Francisco, CA 94158 USA
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  • For correspondence: bo.huang@ucsf.edu
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Abstract

Recent advances in genome engineering have expanded our capabilities to study proteins in their natural states. In particular, the ease and scalability of knocking-in small peptide tags has enabled high throughput tagging and analysis of endogenous proteins. To improve enrichment capacities and expand the functionality of knock-ins using short tags, we developed the tag-assisted split enzyme complementation (TASEC) approach, which uses two orthogonal small peptide tags and their cognate binders to conditionally drive complementation of a split enzyme upon labeled protein expression. Using this approach, we have engineered and optimized the tag-assisted split HaloTag complementation system (TA-splitHalo) and demonstrated its versatile applications in improving the efficiency of knock-in cell enrichment, detection of protein-protein interaction, and isolation of biallelic gene edited cells through multiplexing.

Competing Interest Statement

A patent application has been filed based on this work.

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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 4.0 International license.
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Posted December 02, 2020.
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Versatile labeling and detection of endogenous proteins using tag-assisted split enzyme complementation
Suraj Makhija, David Brown, Struan Bourke, Yina Wang, Shuqin Zhou, Rachel Rudlaff, Rasmi Cheloor-Kovilakam, Bo Huang
bioRxiv 2020.12.01.407072; doi: https://doi.org/10.1101/2020.12.01.407072
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Versatile labeling and detection of endogenous proteins using tag-assisted split enzyme complementation
Suraj Makhija, David Brown, Struan Bourke, Yina Wang, Shuqin Zhou, Rachel Rudlaff, Rasmi Cheloor-Kovilakam, Bo Huang
bioRxiv 2020.12.01.407072; doi: https://doi.org/10.1101/2020.12.01.407072

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