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Logic-gating the HaloTag system with Conditional-Halo-ligator ‘CHalo’ reagents

2025/09/23 by Philipp Mauker, Carmen Zecha, Lucas Dessen-Weissenhorn +19 · 1 voice · 2 citations
Chemical Engineering · Chemistry · Biochemistry, Genetics and Molecular Biology · #Catalysis and Oxidation Reactions #Inorganic Chemistry and Materials #Chemical Synthesis and Analysis

paper · pdf · doi:10.1101/2025.09.23.676741

Abstract

Abstract HaloTag proteins spontaneously ligate onto any chemical reagent featuring a chloroalkane motif ( CA ). We introduce the conditional CHalo motif, which ligates to HaloTag only after uncaging by light or enzymes. (1) Photo-triggered CHalo fluorogenic reagents allow spatiotemporally-specific labeling; (2) photo-triggered CHalo heterodimerisers can photocontrol protein recruitment; and (3) enzyme-triggered CHalo reagents can durably record diverse enzyme activities, and multiplexing them should allow quantitative ratiometric recording of multiple activities in parallel. CHalo thus permits manifold extensions to the HaloTag technology. Abstract Figure

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