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bioRxiv · 10.1101/2020.12.01.407072

Versatile labeling and detection ofendogenous proteins using tag-assistedsplit enzyme complementation

Makhija, S.·Brown, D.·Bourke, S.·Wang, Y.·Zhou, S.·Rudlaff, R.·Cheloor-Kovilakam, R.·Huang, B.
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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.

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Makhija, S., Brown, D., Bourke, S., Wang, Y., Zhou, S., Rudlaff, R., Cheloor-Kovilakam, R., Huang, B.. 2020-12-02. Versatile labeling and detection ofendogenous proteins using tag-assistedsplit enzyme complementation. https://doi.org/10.1101/2020.12.01.407072

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