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

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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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